TW202509069A - Anti-cd161 antibodies and methods of use thereof - Google Patents

Abstract

The present disclosure provides antibodies and antigen-binding portions thereof that bind CD161, bispecific antibodies and multispecific antibodies comprising the same, nucleic acids and vectors encoding the same, and methods of using the same to treat a subject in need thereof.

Description

Translated fromChinese

抗CD161抗體及其使用方法Anti-CD161 antibodies and methods of use thereof

本發明係關於結合CD161之抗體及其抗原結合部分及使用其等來治療有需要個體之方法。The present invention relates to antibodies and antigen-binding portions thereof that bind CD161 and methods of using the same to treat individuals in need thereof.

人類癌症具有許多遺傳及表觀遺傳改變，從而產生潛在可藉由免疫系統識別之新抗原(Sjoblom等人，Science 2006)314(5797):268-274)。繼承免疫系統(其包括T及B淋巴細胞)具有強大的抗癌潛力，具有廣泛能力及精緻特異性來應對不同的腫瘤抗原。免疫療法在癌症治療中之使用係基於腫瘤藉由被識別為自我及非自我來逃避內源免疫反應之前提。腫瘤可使用不同機制逃脫免疫監視且發展出免疫抗性。癌症免疫療法中之新穎方法係抵消此等抗性機制，從而允許內源免疫系統排斥腫瘤。最近免疫調節劑在患有難治性實體腫瘤的患者中之成功提供了免疫系統活化作為免疫治療方式(諸如抗PD-1及抗CTLA-4治療)之功效的概念證明。Human cancers harbor many genetic and epigenetic alterations that generate new antigens that are potentially recognizable by the immune system (Sjoblom et al., Science 2006) 314(5797):268-274). The inherited immune system, which includes T and B lymphocytes, has a powerful anti-cancer potential, with broad capabilities and exquisite specificity to respond to different tumor antigens. The use of immunotherapy in cancer treatment is based on the premise that tumors evade endogenous immune responses by being recognized as self and non-self. Tumors can use different mechanisms to escape immune surveillance and develop immune resistance. Novel approaches in cancer immunotherapy are to counteract these resistance mechanisms, thereby allowing the endogenous immune system to reject the tumor. Recent success with immunomodulatory agents in patients with refractory solid tumors provides proof of concept for the efficacy of immune system activation as an immunotherapy modality, such as anti-PD-1 and anti-CTLA-4 therapy.

儘管免疫療法取得彼等進展，但並非所有腫瘤對經核准之免疫調節劑具有反應(Filley等人，2017)。另外，腫瘤產生抑制性微環境以逃避且抑制免疫反應，此導致基於獨特患者特徵之抗癌劑之有效性之變化(Topalian等人，2014)。對目前免疫療法之免疫抗性仍係一項重大挑戰。因此，需要可在靶向癌症及其他惡性病之免疫抑制環境中誘導免疫反應之另外療法。與放射、化學療法及/或外科手術之組合免疫療法可為癌症提供有效的治療手段。新穎可藥用靶及治療組合將在擴大免疫療法在治療各種癌症中的視野方面發揮關鍵作用。Despite the advances in immunotherapy, not all tumors are responsive to approved immunomodulatory agents (Filley et al., 2017). In addition, tumors create suppressive microenvironments to evade and suppress immune responses, which results in variability in the effectiveness of anticancer agents based on unique patient characteristics (Topalian et al., 2014). Immune resistance to current immunotherapies remains a major challenge. Therefore, there is a need for additional therapies that can induce immune responses in the immunosuppressive environment of targeted cancer and other malignancies. Combination immunotherapy with radiation, chemotherapy, and/or surgery may provide an effective treatment for cancer. New druggable targets and therapeutic combinations will play a key role in expanding the horizons of immunotherapy in the treatment of various cancers.

本發明之一些態樣係關於一種特異性結合CD161之抗體或其抗原結合部分，其包含重鏈可變區(VH)及輕鏈可變區(VL)；其中該VH包含VH互補決定區1 (VH-CDR1)、VH-CDR2及VH-CDR3；其中該VL包含VL-CDR1、VL-CDR2及VL-CDR3；且其中該VH-CDR3包含選自SEQ ID NO: 3、13、23、33、43、53、63、73、83、93、103、113、123、133、143、153、163、173、183、193、203、213、223、233、243、253、263、273、283、293、303、313、323及333所示序列之胺基酸序列。Some aspects of the present invention relate to an antibody or an antigen-binding portion thereof that specifically binds to CD161, comprising a heavy chain variable region (VH) and a light chain variable region (VL); wherein the VH comprises a VH complement determining region 1 (VH-CDR1), a VH-CDR2, and a VH-CDR3; wherein the VL comprises a VL-CDR1, a VL-CDR2, and a VL-CDR3; and wherein the VH-CDR3 comprises a sequence selected from SEQ ID NO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, 133, 143, 153, 163, 173, 183, 193, 203, 213, 223, 233, 243, 253, 263, 273, 283, 293, 303, 313, 323 and the amino acid sequence of the sequence shown in 333.

在一些態樣中，該VH-CDR2包含選自SEQ ID NO: 2、12、22、32、42、52、62、72、82、92、102、112、122、132、142、152、162、172、182、192、202、212、222、232、242、252、262、272、282、292、302、312、322及332所示序列之胺基酸序列。在一些態樣中，該VH-CDR1包含選自SEQ ID NO: 1、11、21、31、41、51、61、71、81、91、101、111、121、131、141、151、161、171、181、191、201、211、221、231、241、251、261、271、281、291、301、311、321及331所示序列之胺基酸序列。在一些態樣中，該VL-CDR3包含選自SEQ ID NO: 6、16、26、36、46、56、66、76、86、96、106、116、126、136、146、156、166、176、186、196、206、216、226、236、246、256、266、276、286、296、306、316、326及336所示序列之胺基酸序列。在一些態樣中，該VL-CDR2包含選自SEQ ID NO: 5、15、25、35、45、55、65、75、85、95、105、115、125、135、145、155、165、175、185、195、205、215、225、235、245、255、265、275、285、295、305、315、325及335所示序列之胺基酸序列。在一些態樣中，該VL-CDR1包含選自SEQ ID NO: 4、14、24、34、44、54、64、74、84、94、104、114、124、134、144、154、164、174、184、194、204、214、224、234、244、254、264、274、284、294、304、314、324及334所示序列之胺基酸序列。In some aspects, the VH-CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, and 332. In some aspects, the VH-CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, and 331. In some aspects, the VL-CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, and 336. In some aspects, the VL-CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325 and 335. In some aspects, the VL-CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, and 334.

在一些態樣中，該抗體或其抗原結合部分包含：(i)包含SEQ ID NO: 1所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 2所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 3所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 4所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 5所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 6所示胺基酸序列之VL-CDR3；(ii)包含SEQ ID NO: 91所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 92所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 93所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 94所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 95所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 96所示胺基酸序列之VL-CDR3；(iii)包含SEQ ID NO: 111所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 112所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 113所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 114所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 115所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 116所示胺基酸序列之VL-CDR3；(iv)包含SEQ ID NO: 141所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 142所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 143所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 144所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 145所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 146所示胺基酸序列之VL-CDR3；(v)包含SEQ ID NO: 211所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 212所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 213所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 214所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 215所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 216所示胺基酸序列之VL-CDR3；(vi)包含SEQ ID NO: 221所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 222所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 223所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 224所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 225所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 226所示胺基酸序列之VL-CDR3；(vii)包含SEQ ID NO: 271所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 272所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 273所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 274所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 275所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 276所示胺基酸序列之VL-CDR3；(viii)包含SEQ ID NO: 281所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 282所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 283所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 284所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 285所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 286所示胺基酸序列之VL-CDR3；(ix)包含SEQ ID NO: 291所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 292所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 293所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 294所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 295所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 296所示胺基酸序列之VL-CDR3；(x)包含SEQ ID NO: 301所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 302所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 303所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 304所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 305所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 306所示胺基酸序列之VL-CDR3；或(xi) (i)至(x)之任何組合。In some aspects, the antibody or antigen-binding portion thereof comprises: (i) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 6; (ii) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 91, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 92, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 93, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 94, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 95, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 96, a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: (iii) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 111, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 112, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 113, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 114, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 116; (iv) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 144, NO: 144, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 145, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 146, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 147; (v) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 213, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 216; (vi) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 221, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: (vii) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 271, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 272, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 273, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 274, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 275, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 276; (viii) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 281, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 282, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 283, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 284, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 285, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 286; (ix) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 291, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 292, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 293, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: (x) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 302, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 303, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 306; or (xi) any combination of (i) to (x).

在一些態樣中，該VH包含與選自SEQ ID NO: 7、17、27、37、47、57、67、77、87、97、107、117、127、137、147、157、167、177、187、197、207、217、227、237、247、257、267、277、287、297、707、317、327及337之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列。在一些態樣中，該VL包含與選自SEQ ID NO: 8、18、28、38、48、58、68、78、88、98、108、118、128、138、148、158、168、178、188、198、208、218、228、238、248、258、268、278、288、298、308、318、328及338之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列。In some aspects, the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337. In some aspects, the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.

本發明之一些態樣係關於特異性結合CD161之抗體或其抗原結合部分，其包含可變重(VH)域及可變輕(VL)域，其中該VH包含與選自SEQ ID NO: 7、17、27、37、47、57、67、77、87、97、107、117、127、137、147、157、167、177、187、197、207、217、227、237、247、257、267、277、287、297、707、317、327及337之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且其中該VL包含與選自SEQ ID NO: 8、18、28、38、48、58、68、78、88、98、108、118、128、138、148、158、168、178、188、198、208、218、228、238、248、258、268、278、288、298、308、318、328及338之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列。Some aspects of the present invention relate to antibodies or antigen-binding portions thereof that specifically bind to CD161, comprising a variable heavy (VH) domain and a variable light (VL) domain, wherein the VH comprises a polypeptide selected from SEQ ID NO: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337 have an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity; and wherein the VL comprises an amino acid sequence selected from SEQ ID NO: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328 and 338 have an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity.

在一些態樣中，該VH包含選自SEQ ID NO: 7、17、27、37、47、57、67、77、87、97、107、117、127、137、147、157、167、177、187、197、207、217、227、237、247、257、267、277、287、297、707、317、327及337之胺基酸序列。在一些態樣中，該VL包含選自SEQ ID NO: 8、18、28、38、48、58、68、78、88、98、108、118、128、138、148、158、168、178、188、198、208、218、228、238、248、258、268、278、288、298、308、318、328及338之胺基酸序列。In some aspects, the VH comprises an amino acid sequence selected from SEQ ID NO: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337. In some aspects, the VL comprises an amino acid sequence selected from SEQ ID NO: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.

在一些態樣中，(i)該VH包含與選自SEQ ID NO: 7之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 8之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(ii)該VH包含與選自SEQ ID NO: 97之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 98之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(iii)該VH包含與選自SEQ ID NO: 117之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 118之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(iv)該VH包含與選自SEQ ID NO: 147之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 148之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(v)該VH包含與選自SEQ ID NO: 217之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 218之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(vi)該VH包含與選自SEQ ID NO: 227之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 228之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(vii)該VH包含與選自SEQ ID NO: 277之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 278之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(viii)該VH包含與選自SEQ ID NO: 287之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 288之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(ix)該VH包含與選自SEQ ID NO: 297之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 298之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；(x)該VH包含與選自SEQ ID NO: 307之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 308之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；或(xi) (i)至(x)之任何組合。In some aspects, (i) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 7; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 8; (ii) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 97; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 98 has at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 117; (iii) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 117; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 118; (iv) the VH comprises an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: wherein the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 148; and (v) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 217; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 148. 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 218; (vi) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 227; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 228; (vii) the VH comprises an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: wherein the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 278; and (viii) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 287; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 278. 288 has at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 297; (ix) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 297; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 298; (x) the VH comprises an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: The invention further comprises an amino acid sequence selected from SEQ ID NO: 307 having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity; and the VL comprises an amino acid sequence selected from SEQ ID NO: 308 having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity.

在一些態樣中，該抗體或其抗原結合部分含有：(i)包含SEQ ID NO: 7所示胺基酸序列之VH、及包含SEQ ID NO: 8所示胺基酸序列之VL；(ii)包含SEQ ID NO: 97所示胺基酸序列之VH、及包含SEQ ID NO: 98所示胺基酸序列之VL；(iii)包含SEQ ID NO: 117所示胺基酸序列之VH、及包含SEQ ID NO: 118所示胺基酸序列之VL；(iv)包含SEQ ID NO: 147所示胺基酸序列之VH、及包含SEQ ID NO: 148所示胺基酸序列之VL；(v)包含SEQ ID NO: 217所示胺基酸序列之VH、及包含SEQ ID NO: 218所示胺基酸序列之VL；(vi)包含SEQ ID NO: 227所示胺基酸序列之VH、及包含SEQ ID NO: 228所示胺基酸序列之VL；(vii)包含SEQ ID NO: 277所示胺基酸序列之VH、及包含SEQ ID NO: 278所示胺基酸序列之VL；(viii)包含SEQ ID NO: 287所示胺基酸序列之VH、及包含SEQ ID NO: 288所示胺基酸序列之VL；(ix)包含SEQ ID NO: 297所示胺基酸序列之VH、及包含SEQ ID NO: 298所示胺基酸序列之VL；(x)包含SEQ ID NO: 307所示胺基酸序列之VH、及包含SEQ ID NO: 308所示胺基酸序列之VL；或(xi) (i)至(x)之任何組合。本發明之一些態樣係關於結合與本文揭示之抗體或其抗原結合部分相同的抗原決定基之抗體或其抗原結合部分。本發明之一些態樣係關於與本文揭示之抗體或其抗原結合部分交叉競爭結合CD161之抗體或其抗原結合部分。In some aspects, the antibody or its antigen-binding portion comprises: (i) a VH comprising the amino acid sequence of SEQ ID NO: 7, and a VL comprising the amino acid sequence of SEQ ID NO: 8; (ii) a VH comprising the amino acid sequence of SEQ ID NO: 97, and a VL comprising the amino acid sequence of SEQ ID NO: 98; (iii) a VH comprising the amino acid sequence of SEQ ID NO: 117, and a VL comprising the amino acid sequence of SEQ ID NO: 118; (iv) a VH comprising the amino acid sequence of SEQ ID NO: 147, and a VL comprising the amino acid sequence of SEQ ID NO: 148; (v) a VH comprising the amino acid sequence of SEQ ID NO: 217, and a VL comprising the amino acid sequence of SEQ ID NO: 218; (vi) a VH comprising the amino acid sequence of SEQ ID NO: 227, and a VL comprising the amino acid sequence of SEQ ID NO: (vii) a VH comprising the amino acid sequence of SEQ ID NO: 277, and a VL comprising the amino acid sequence of SEQ ID NO: 278; (viii) a VH comprising the amino acid sequence of SEQ ID NO: 287, and a VL comprising the amino acid sequence of SEQ ID NO: 288; (ix) a VH comprising the amino acid sequence of SEQ ID NO: 297, and a VL comprising the amino acid sequence of SEQ ID NO: 298; (x) a VH comprising the amino acid sequence of SEQ ID NO: 307, and a VL comprising the amino acid sequence of SEQ ID NO: 308; or (xi) any combination of (i) to (x). Some aspects of the invention relate to antibodies or antigen-binding portions thereof that bind to the same antigenic determinant as the antibodies or antigen-binding portions thereof disclosed herein. Some aspects of the invention relate to antibodies, or antigen-binding portions thereof, that cross-compete with the antibodies, or antigen-binding portions thereof, disclosed herein for binding to CD161.

在一些態樣中，該抗體或其抗原結合部分以小於約1000 nM、小於約500 nM、小於約100 nM、小於約50 nM或小於約10 nM之K_D結合CD161。在一些態樣中，該抗體或其抗原結合部分以小於約50 nM之K_D結合CD161。在一些態樣中，該抗體或其抗原結合部分以小於約10 nM之K_D結合CD161。In some aspects, the antibody, or antigen-binding portion thereof, binds CD161 with a_KD of less than about 1000 nM, less than about 500 nM, less than about 100 nM, less than about 50 nM, or less than about 10 nM. In some aspects, the antibody, or antigen-binding portion thereof, binds CD161 with a_KD of less than about 50 nM. In some aspects, the antibody, or antigen-binding portion thereof, binds CD161 with a_KD of less than about 10 nM.

在一些態樣中，該抗體或其抗原結合部分抑制CD161與C型凝集素域家族2成員D (CLEC2D)之間的相互作用。In some aspects, the antibody, or antigen-binding portion thereof, inhibits the interaction between CD161 and C-type lectin domain family 2, member D (CLEC2D).

在一些態樣中，該抗體或其抗原結合部分能夠誘導或增強免疫細胞產生一或多種細胞激素。在一些態樣中，該一或多種細胞激素包含IL2、TNFa、IFNg或其任何組合。In some aspects, the antibody or its antigen-binding portion can induce or enhance immune cells to produce one or more cytokines. In some aspects, the one or more cytokines include IL2, TNFa, IFNg or any combination thereof.

在一些態樣中，該抗體之該抗原結合部分包含VHH、vNAR、微體(microbody)、奈米抗體(nanobody)、scFv或其任何組合。In some aspects, the antigen-binding portion of the antibody comprises a VHH, a vNAR, a microbody, a nanobody, a scFv, or any combination thereof.

本發明之一些態樣係關於包含本文揭示之抗體或其抗原結合部分之多特異性抗體。本發明之一些態樣係關於包含本文揭示之抗體或其抗原結合部分之雙特異性抗體。Some aspects of the invention relate to multispecific antibodies comprising an antibody disclosed herein or an antigen-binding portion thereof. Some aspects of the invention relate to bispecific antibodies comprising an antibody disclosed herein or an antigen-binding portion thereof.

本發明之一些態樣係關於編碼本文揭示之抗體或其抗原結合部分、本文揭示之多特異性抗體、或本文揭示之雙特異性抗體之核酸分子或一組核酸分子。Some aspects of the invention pertain to a nucleic acid molecule or a set of nucleic acid molecules encoding an antibody disclosed herein or an antigen-binding portion thereof, a multispecific antibody disclosed herein, or a bispecific antibody disclosed herein.

本發明之一些態樣係關於一種載體或一組載體，其包含本文揭示的核酸分子或一組核酸分子。在一些態樣中，該載體係病毒載體。Some aspects of the present invention relate to a vector or a group of vectors, which comprises a nucleic acid molecule or a group of nucleic acid molecules disclosed herein. In some aspects, the vector is a viral vector.

本發明之一些態樣係關於包含本文揭示之核酸分子或一組核酸分子或本文揭示之載體或一組載體之宿主細胞。Some aspects of the invention relate to host cells comprising a nucleic acid molecule or a set of nucleic acid molecules disclosed herein or a vector or a set of vectors disclosed herein.

本發明之一些態樣係關於包含本文揭示之抗體或其抗原結合部分、本文揭示之多特異性抗體、本文揭示之雙特異性抗體、本文揭示之核酸分子或一組核酸分子、本文揭示之載體或一組載體、或本文揭示之宿主細胞及醫藥上可接受之載劑之醫藥組合物。Some aspects of the invention relate to pharmaceutical compositions comprising an antibody or antigen-binding portion thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or a set of nucleic acid molecules disclosed herein, a vector or a set of vectors disclosed herein, or a host cell disclosed herein and a pharmaceutically acceptable carrier.

本發明之一些態樣係關於一種治療有需要個體之疾病或病症之方法，其包括對該個體投與本文揭示之抗體或其抗原結合部分、本文揭示之多特異性抗體、本文揭示之雙特異性抗體、本文揭示之核酸分子或一組核酸分子、本文揭示之載體或一組載體、本文揭示之宿主細胞或本文揭示之醫藥組合物。在一些態樣中，該疾病或病症包含癌症。Some aspects of the invention relate to a method of treating a disease or condition in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or a set of nucleic acid molecules disclosed herein, a vector or a set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein. In some aspects, the disease or condition comprises cancer.

本發明之一些態樣係關於一種誘導有需要個體中之免疫反應之方法，其包括對該個體投與本文揭示之抗體或其抗原結合部分、本文揭示之多特異性抗體、本文揭示之雙特異性抗體、本文揭示之核酸分子或一組核酸分子、本文揭示之載體或一組載體、本文揭示之宿主細胞或本文揭示之醫藥組合物。在一些態樣中，該個體罹患癌症。Some aspects of the invention relate to a method of inducing an immune response in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or a set of nucleic acid molecules disclosed herein, a vector or a set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein. In some aspects, the subject suffers from cancer.

本發明之一些態樣係關於一種治療有需要個體之癌症之方法，其包括對該個體投與本文揭示之抗體或其抗原結合部分、本文揭示之多特異性抗體、本文揭示之雙特異性抗體、本文揭示之核酸分子或一組核酸分子、本文揭示之載體或一組載體、本文揭示之宿主細胞或本文揭示之醫藥組合物。Some aspects of the invention relate to a method of treating cancer in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or a set of nucleic acid molecules disclosed herein, a vector or a set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein.

在一些態樣中，該癌症選自聽神經瘤、急性淋巴球性白血病、急性淋巴球性白血病、腺癌、及泌尿系統癌、及上皮細胞癌、血管肉瘤、星形細胞瘤、基底細胞癌、膽管癌、膽道癌、膀胱癌、骨癌、腦癌、腦幹神經膠質瘤、乳癌、支氣管癌、伯奇氏淋巴瘤(Burkitt's lymphoma)及邊緣區B細胞淋巴瘤、腎上腺癌、肛門區癌、消化系統癌、內分泌系統癌、食道癌、副甲狀腺癌、陰莖癌、呼吸系统癌、小腸癌、輸尿管癌、尿道癌、子宮頸癌、子宮內膜癌、輸卵管癌、腎盂癌、陰道癌、外陰癌、中樞神經系統(CNS)癌症、子宮頸癌、軟骨肉瘤、脊索瘤、絨毛膜癌、慢性白血病、慢性淋巴球性白血病、慢性髓細胞性(粒細胞性)白血病、結腸癌、結腸肉瘤、結腸直腸癌、結締組織癌、顱咽管瘤、囊腺癌、胚胎癌、子宮內膜癌、內皮肉瘤、環境誘導型癌症(包括彼等由石棉誘導者)、室管膜瘤、表皮樣癌、上皮樣癌、食道癌、食道上皮細胞癌、尤因氏肉瘤(Ewing's tumor)、眼癌、纖維肉瘤、胃癌(gastric cancer)、胃腸癌、生殖細胞腫瘤、神經膠質母細胞瘤(例如多形性神經膠質母細胞瘤)、神經膠質瘤、頭頸癌、重鏈病(heavy chain disease)、血管母細胞瘤、肝癌瘤、霍奇金氏病(Hodgkin's disease)、上皮內贅瘤、卡波西氏肉瘤(Kaposi's sarcoma)、腎癌(例如腎細胞癌(RCC))、喉頭癌、平滑肌肉瘤、白血病、脂肪肉瘤、肝癌、肺癌(lung cancer) (小細胞、大細胞)、肺癌(lung carcinoma)、淋巴管內皮肉瘤、淋巴管肉瘤、被套細胞淋巴瘤、髓質癌、神經管胚細胞瘤、黑色素瘤、腦膜瘤、間皮瘤、多發性骨髓瘤、骨髓母細胞前髓細胞骨髓單核球性單核細胞紅血球性白血病、黏液肉瘤、鼻咽癌、中樞神經系統(CNS)贅瘤、神經母細胞瘤、非霍奇金氏病(non-Hodgkin's disease)、非小細胞肺癌(non-small cell lung cancer；NSCLC)、非小細胞肺上皮細胞癌(non-small cell lung carcinoma)、寡樹突神經膠質瘤、口腔癌(例如唇、舌、口及咽)、成骨肉瘤、骨肉瘤、卵巢癌、胰臟癌、乳突腺癌、乳突癌、小兒肉瘤、松果腺瘤(pinealoma)、垂體腺瘤、真性紅血球增多症(Polycythemia vera)、淋巴瘤、原發性CNS淋巴瘤、前列腺癌(例如激素難治性前列腺腺癌)、直腸癌、腎癌(例如透明細胞癌)、視網膜母細胞瘤、橫紋肌肉瘤、肉瘤、軟組織肉瘤、皮脂腺癌、精細胞瘤、鼻腔鼻竇自然殺手(sinonasal natural killer)、皮膚癌、小細胞肺癌(SCLC)、兒童實體腫瘤、脊髓軸瘤、鱗狀細胞癌(squamous cell cancer)、鱗狀細胞癌(squamous cell carcinoma)、胃癌(stomach cancer)、汗腺癌、滑膜瘤、睾丸癌、甲狀腺癌、腫瘤血管生成、子宫癌、病毒相關癌症或病毒起源之癌症(例如人類乳頭瘤病毒(HPV相關或來源腫瘤))、華氏巨球蛋白血症(Waldenstrom's macroglobulinemia)及威爾姆氏腫瘤(Wilm's tumor)；及該等癌症之任何組合。In some aspects, the cancer is selected from acoustic neuroma, acute lymphocytic leukemia, acute lymphocytic leukemia, adenocarcinoma, and urinary system cancer, and epithelial cell carcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct carcinoma, bile duct cancer, bladder cancer, bone cancer, brain cancer, brain stem neurofibroma, breast cancer, bronchial carcinoma, Birch's lymphoma (Burkitt's lymphoma) and marginal B-cell lymphoma, adrenal cancer, anal cancer, digestive system cancer, endocrine system cancer, esophageal cancer, parathyroid cancer, penile cancer, respiratory system cancer, small intestine cancer, ureteral cancer, urethral cancer, cervical cancer, endometrial cancer, fallopian tube cancer, renal pelvis cancer, vaginal cancer, vulvar cancer, central nervous system (CNS) cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, Chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, colon cancer, colon sarcoma, colorectal cancer, connective tissue cancer, cranio-pharyngioma, cystadenocarcinoma, embryonal carcinoma, endometrial carcinoma, endothelial sarcoma, environmentally induced cancers (including those induced by asbestos), ependymoma, epidermoid carcinoma, epithelioid carcinoma, esophageal cancer, esophageal epithelial cell carcinoma, Ewing's sarcoma tumor), eye cancer, fibrosarcoma, gastric cancer, gastrointestinal cancer, germ cell tumor, neuroglioblastoma (e.g., multiforme neuroglioblastoma), neuroglioma, head and neck cancer, heavy chain disease, hemangioblastoma, hepatocarcinoma, Hodgkin's disease, intraepithelial neoplasia, Kaposi's sarcoma, kidney cancer (e.g., renal cell carcinoma (RCC)), laryngeal cancer, leiomyosarcoma, leukemia, liposarcoma, liver cancer, lung cancer (small cell, large cell), lung cancer (lung carcinoma), lymphangioendothelioma, lymphangiosarcoma, mantle cell lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloblastic promyelocytic myelomonocytic monocytic erythroblastic leukemia, myxosarcoma, nasopharyngeal carcinoma, central nervous system (CNS) neoplasms, neuroblastoma, non-Hodgkin's disease, non-small cell lung cancer (NSCLC), non-small cell lung cancer (NSCLC), carcinoma), oligodendrocyte neuroglia, oral cancer (e.g., lip, tongue, mouth, and pharynx), osteogenic sarcoma, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pediatric sarcoma, pinealoma, pituitary adenoma, polycythemia vera, lymphoma, primary CNS lymphoma, prostate cancer (e.g., hormone-refractory prostate adenocarcinoma), rectal cancer, kidney cancer (e.g., clear cell carcinoma), retinoblastoma, rhabdomyosarcoma, sarcoma, Soft tissue sarcoma, sebaceous gland carcinoma, spermatoma, sinonasal natural killer, skin cancer, small cell lung cancer (SCLC), solid tumors in children, spinal cord axonal tumors, squamous cell carcinoma cancer), squamous cell carcinoma, stomach cancer, sweat gland carcinoma, synovium, testicular cancer, thyroid cancer, tumor angiogenesis, uterine cancer, virus-related cancers or cancers of viral origin (e.g., human papillomavirus (HPV-related or derived tumors)), Waldenstrom's macroglobulinemia, and Wilm's tumor; and any combination of these cancers.

本發明之一些態樣係關於一種治療有需要個體之傳染病之方法，其包括對該個體投與本文揭示之抗體或其抗原結合部分、本文揭示之多特異性抗體、本文揭示之雙特異性抗體、本文揭示之核酸分子或一組核酸分子、本文揭示之載體或一組載體、本文揭示之宿主細胞或本文揭示之醫藥組合物。在一些態樣中，該傳染病包含：(i)流行性感冒、皰疹、梨形鞭毛蟲屬(Giardia)、瘧疾、利什曼原蟲(Leishmania)或其任何組合之感染；(ii)人類免疫缺陷病毒(HIV)、肝炎病毒、皰疹病毒、腺病毒、流行性感冒病毒、黃病毒科、伊科病毒(echovirus)、鼻病毒、柯薩奇病毒(coxsackie virus)、冠狀病毒、呼吸道融合病毒、腮腺炎病毒、輪狀病毒(rotavirus)、麻疹病毒、蕁麻疹病毒、小病毒、牛痘病毒、HTLV病毒、登革熱病毒(dengue virus)、乳頭瘤病毒、軟疣病毒、脊髓灰質炎病毒、狂犬病病毒、JC病毒或蟲媒病毒腦炎病毒或其任何組合之感染；(iii)披衣菌(chlamydia)、立克次體細菌(rickettsial bacteria)、分枝桿菌、葡萄球菌、鏈球菌、肺炎球菌、腦膜炎雙球菌、淋球菌、克雷伯氏菌(klebsiella)、變形桿菌屬、沙雷氏菌(serratia)、假單胞菌、軍團菌屬(legionella)、白喉桿菌(diphtheria)、沙門氏菌(salmonella)、桿菌、霍亂、破傷風、肉毒症、炭疽、瘟疫、鉤端螺旋體病、及萊姆氏病(Lyme’s disease)細菌、或其任何組合之感染；(iv)念珠菌屬(Candida)、新型隱球菌(Cryptococcus neoformans)、麴菌屬(Aspergillus)、毛黴目(Mucorales)屬、申克氏胞絲菌(Sporothrix schenkii)、皮炎芽生菌(Blastomyces dermatitidis)、巴西副球孢子菌(Paracoccidioides brasiliensis)、粗球黴菌(Coccidioides immitis)、或莢膜組織漿菌(Histoplasma capsulatum)、或其任何組合之感染；(v)溶組織內阿米巴(Entamoeba histolytica)、結腸小袋纖毛蟲(Balantidium coli)、福氏耐格里阿米巴原蟲(Naegleriafowleri)、棘阿米巴科(Acanthamoeba sp.)、蘭氏賈第鞭毛蟲(Giardia lambia)、隱孢子蟲(Cryptosporidium sp.)、卡式肺囊蟲(Pneumocystis carinii)、間日瘧原蟲(Plasmodium vivax)、巴貝斯原蟲(Babesia microti)、布氏錐蟲(Trypanosoma brucei)、克氏錐蟲(Trypanosoma cruzi)、杜氏利什曼原蟲(Leishmania donovani)、岡地弓漿蟲病(Toxoplasma gondi)、或巴西日圓線蟲(Nippostrongylus brasiliensis)、或其任何組合之感染；或(vi) (i)至(v)之任何組合。Some aspects of the invention relate to a method of treating an infectious disease in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or a set of nucleic acid molecules disclosed herein, a vector or a set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein. In some aspects, the infectious disease comprises: (i) infection with influenza, herpes, Giardia, malaria, Leishmania, or any combination thereof; (ii) infection with human immunodeficiency virus (HIV), hepatitis virus, herpes virus, adenovirus, influenza virus, Flaviviridae, echovirus, rhinovirus, coxsackie virus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum virus, polio virus, rabies virus, JC virus, or encephalitis virus, or any combination thereof; (iii) infection withchlamydia ,rickettsial bacteria , (iv) infections caused by Candida,Cryptococcusneoformans ,Aspergillus ,Mucorales ,Sporothrix schenkii,andany combinationthereof. (v) infection with Entamoebahistolytica ,Balantidium coli , Naegleria fowleri,Acanthamoebasp .,Giardia lambia,Cryptosporidiumsp.,Pneumocystiscarinii ,Plasmodium vivax, or any combination thereof; ),Babesia microti ,Trypanosoma brucei ,Trypanosoma cruzi ,Leishmania donovani , Toxoplasma gondi, or Nippostrongylus brasiliensis, or any combination thereof; or (vi) any combination of (i) to (v).

本發明之一些態樣係關於一種治療有需要個體之自體免疫疾病之方法，其包括對該個體投與本文揭示之抗體或其抗原結合部分、本文揭示之多特異性抗體、本文揭示之雙特異性抗體、本文揭示之核酸分子或一組核酸分子、本文揭示之載體或一組載體、本文揭示之宿主細胞或本文揭示之醫藥組合物。Some aspects of the invention relate to a method of treating an autoimmune disease in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding portion thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or a set of nucleic acid molecules disclosed herein, a vector or a set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein.

本發明之一些態樣係關於一種活化免疫細胞之方法，其包括使該免疫細胞與本文揭示之抗體或其抗原結合部分、本文揭示之多特異性抗體、本文揭示之雙特異性抗體、本文揭示之核酸分子或一組核酸分子、本文揭示之載體或一組載體、本文揭示之宿主細胞或本文揭示之醫藥組合物接觸。Some aspects of the invention relate to a method of activating an immune cell, comprising contacting the immune cell with an antibody or antigen-binding portion thereof disclosed herein, a multispecific antibody disclosed herein, a bispecific antibody disclosed herein, a nucleic acid molecule or a set of nucleic acid molecules disclosed herein, a vector or a set of vectors disclosed herein, a host cell disclosed herein, or a pharmaceutical composition disclosed herein.

相關申請案之交叉參考Cross-reference to related applications

本PCT申請案主張2023年5月4日申請之美國臨時申請案第63/500,122號之優先權，該案以全文引用之方式併入本文中。 以電子方式提交之序列表之參考This PCT application claims priority to U.S. Provisional Application No. 63/500,122 filed on May 4, 2023, which is incorporated herein by reference in its entirety.Reference to Sequence Listing Submitted Electronically

序列表之內容以電子方式提交(名稱：5037_001PC01_Seqlisting_ST26；大小：304,752位元组；且創建日期：2024年5月3日)，且該申請案之全文以引用之方式併入本文中。The contents of the sequence listing are submitted electronically (name: 5037_001PC01_Seqlisting_ST26; size: 304,752 bytes; and creation date: May 3, 2024), and the entire text of the application is incorporated herein by reference.

本發明係關於特異性結合CD161之抗體(在本文中稱為抗CD161抗體)及其抗原結合部分。本發明之一些態樣係關於包含該抗CD161抗體之雙特異性或多特異性抗體。本發明之其他態樣係關於治療有需要個體之疾病或病症之方法，其包括對該個體投與本文描述之抗體或其抗原結合部分。The present invention relates to antibodies that specifically bind to CD161 (referred to herein as anti-CD161 antibodies) and antigen-binding portions thereof. Some aspects of the present invention relate to bispecific or multispecific antibodies comprising the anti-CD161 antibodies. Other aspects of the present invention relate to methods for treating a disease or disorder in an individual in need thereof, comprising administering to the individual an antibody described herein or an antigen-binding portion thereof.

在更詳細描述本發明之前，應理解，本發明不限於描述之特定組合物或製程步驟，其當然變化。如熟習此項技術者在閱讀本發明後所明瞭，本文中描述及繪示之各個別態樣具有離散組分及特徵，其在不脫離本發明之範疇或精神下可容易地自任何其他幾個態樣之特徵分離或與任何其他幾個態樣之特徵組合。任何列舉的方法可以所列舉的事件之順序或以邏輯上可能的任一其他順序進行。Before describing the present invention in more detail, it should be understood that the present invention is not limited to the specific compositions or process steps described, which may of course vary. As will be apparent to one skilled in the art upon reading the present invention, each individual aspect described and illustrated herein has discrete components and features that may be readily separated from or combined with the features of any other aspects without departing from the scope or spirit of the present invention. Any enumerated method may be performed in the order of events enumerated or in any other order that is logically possible.

本文所提供的標題不是本發明之各種態樣之限制，其可總體上引用說明書來定義。亦應理解，本文所用的術語係出於僅描述特定態樣之目的而不欲具限制性。 I. 術語The headings provided herein are not limitations of the various aspects of the invention, which may be generally defined by reference to the specification. It should also be understood that the terms used herein are for the purpose of describing specific aspects only and are not intended to be limiting.I. Terminology

為了可更輕易地明瞭本發明，首先定義某些術語。在整個詳細描述中闡述另外定義。In order to more easily understand the present invention, certain terms are first defined. Additional definitions are set forth throughout the detailed description.

應注意，術語「一」或「一個」實體係指一或多個該實體；例如，「一核苷酸序列」應理解為表示一或多個核苷酸序列。因此，術語「一」(或「一個」)、「一或多個」、及「至少一個」可在本文中互換使用。It should be noted that the term "a" or "an" entity refers to one or more of the entity; for example, "a nucleotide sequence" should be understood to mean one or more nucleotide sequences. Therefore, the terms "a" (or "one"), "one or more", and "at least one" can be used interchangeably herein.

此外，「及/或」在本文中使用的地方將被視為特定揭示兩個特定特徵或組分中之各者，連同或不連同另一者。因此，術語「及/或」如在本文片語諸如「A及/或B」中所用意欲包括「A及B」、「A或B」、「A」 (僅)及「B」 (僅)。同樣地，術語「及/或」如在片語諸如「A、B及/或C」中所用意欲涵蓋以下態樣中之各者：A、B及C；A、B或C；A或C；A或B；B或C；A及C；A及B；B及C；A(僅)；B (僅)；及C (僅)。In addition, where "and/or" is used herein, it will be considered to specifically disclose each of the two specified features or components, with or without the other. Therefore, the term "and/or" as used in phrases such as "A and/or B" herein is intended to include "A and B", "A or B", "A" (only) and "B" (only). Similarly, the term "and/or" as used in phrases such as "A, B and/or C" is intended to cover each of the following: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (only); B (only); and C (only).

應理解，在本文中任何處以語言「包含」描述態樣時，亦提供以「由......組成」及/或「基本上由......組成」描述之其他類似態樣。It should be understood that, where the language “comprising” is used anywhere in this document to describe an aspect, other similar aspects described with “consisting of” and/or “consisting essentially of” are also provided.

除非另有定義，否則本文所使用的所有技術及科學術語具有與熟習本發明所屬技術者通常所理解相同的含義。例如，Concise Dictionary of Biomedicine and Molecular Biology，Juo, Pei-Show，第2版，2002年，CRC Press；The Dictionary of Cell and Molecular Biology，第3版，1999年，Academic Press；及Oxford Dictionary Of Biochemistry And Molecular Biology，修訂版，2000年，Oxford University Press提供熟習此項技術者本發明中所使用之許多術語之一般辭典。Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd edition, 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd edition, 1999, Academic Press; and Oxford Dictionary Of Biochemistry And Molecular Biology, Revised Edition, 2000, Oxford University Press provide general dictionaries of many of the terms used in the present invention for those skilled in the art.

單位、前綴、及符號係以其國際單位制(Système International de Unites；SI)接受之形式表示。數字範圍包含界定範圍之數字。除非另作指明，否則核苷酸序列以5’至3’方向從左至右書寫。胺基酸序列以胺基至羧基方向從左至右書寫。本文所提供的標題不是本發明之各種態樣之限制，其可總體上引用說明書來進行。因此，緊接在下文中定義之術語係以其全文引用本說明書的方式更全面地定義。Units, prefixes, and symbols are expressed in the form accepted by the International System of Units (Système International de Unites; SI). Numerical ranges include numbers that define the range. Unless otherwise specified, nucleotide sequences are written from left to right in the 5' to 3' direction. Amino acid sequences are written from left to right in the amine to carboxyl direction. The titles provided herein are not limitations of the various aspects of the present invention, which can be generally cited in the specification. Therefore, the terms defined immediately below are more fully defined by citing the specification in its entirety.

術語「約」在本文中用於意指近似(approximately)、約(roughly)、約(around)、或在......之區域中。當術語「約」與數值範圍結合使用時，其藉由擴展所述數值之上及之下的邊界來修飾該範圍。一般而言，術語「約」可修飾數值高於或低於規定值，例如，向上或向下(高或低) 10%之差異。The term "about" is used herein to mean approximately, roughly, around, or in the region of. When the term "about" is used in conjunction with a numerical range, it modifies the range by extending the boundaries above and below the numerical values. Generally, the term "about" can modify a numerical value above or below the stated value, for example, up or down (higher or lower) by a 10% difference.

術語「抗體」在一些態樣中係指包含經雙硫鍵互連之至少兩個重(H)鏈及兩個輕(L)鏈之蛋白質或其抗原結合部分。每個重鏈由重鏈可變區(在本文中縮寫為VH)及重鏈恆定區(在本文中縮寫為CH)組成。在一些抗體(例如天然存在之IgG抗體)中，該重鏈恆定區由鉸鏈及三個域CH1、CH2及CH3組成。在一些實施例(例如天然存在之IgG抗體)中，每個輕鏈由輕鏈可變區(在本文中縮寫為VL)及輕鏈恆定區組成。該輕鏈恆定區由一個域(在本文中縮寫為CL)組成。該等VH及VL區可進一步細分為稱為互補決定區(CDR)之超變區，其間散佈著更加保守之稱為框架區(FR)之區域。每個VH及VL由自胺基端至羧基端按以下順序排列之三個CDR及四個FR組成：FR1、CDR1、FR2、CDR2、FR3、CDR3及FR4。該等重鏈及輕鏈之可變區含有與抗原相互作用之結合域。該等抗體之恆定區可介導免疫球蛋白結合至宿主組織或因子，包括免疫系統之各種細胞(例如效應細胞)及典型補體系統之第一組分(C1q)。重鏈可具有或不具有C端離胺酸。除非本文另有指明，否則該等可變區中之胺基酸使用Kabat編號系統進行編號且恆定區中之彼等使用EU系統進行編號。The term "antibody" in some aspects refers to a protein or antigen-binding portion thereof comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region (abbreviated herein as CH). In some antibodies (e.g., naturally occurring IgG antibodies), the heavy chain constant region consists of a hinge and three domains CH1, CH2, and CH3. In some embodiments (e.g., naturally occurring IgG antibodies), each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain (abbreviated herein as CL). The VH and VL regions can be further subdivided into hypervariable regions called complementation determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of the antibodies can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The heavy chain may or may not have a C-terminal lysine. Unless otherwise indicated herein, the amino acids in the variable regions are numbered using the Kabat numbering system and those in the constant regions are numbered using the EU system.

「IgG抗體」例如人類IgG1、IgG2、IgG3及IgG4抗體如本文所用在一些態樣中具有天然存在之IgG抗體之結構，亦即其具有與相同亞類之天然存在之IgG抗體相同數目之重鏈及輕鏈及雙硫鍵。例如，抗CD161 IgG1、IgG2、IgG3或IgG4抗體由兩個重鏈(HC)及兩個輕鏈(LC)組成，其中該兩個HC及LC經天然存在之IgG1、IgG2、IgG3及IgG4抗體中分別出現之相同數目及位置之雙硫橋連接(除非該抗體已經突變以修飾該等雙硫橋)。"IgG antibodies" such as human IgG1, IgG2, IgG3 and IgG4 antibodies as used herein have the structure of naturally occurring IgG antibodies in some aspects, i.e., they have the same number of heavy chains and light chains and disulfide bonds as naturally occurring IgG antibodies of the same subclass. For example, an anti-CD161 IgG1, IgG2, IgG3 or IgG4 antibody is composed of two heavy chains (HC) and two light chains (LC), wherein the two HCs and LCs are connected by disulfide bridges of the same number and position as those found in naturally occurring IgG1, IgG2, IgG3 and IgG4 antibodies, respectively (unless the antibody has been mutated to modify the disulfide bridges).

抗體通常以高親和力特異性結合至其同源抗原，由10^-5至10^-11M或更小之解離常數(K_D)反映。大於約10^-4M之任何K_D一般被認為指示非特異性結合。如本文所用，「特異性結合」至抗原之抗體係指以高親和力結合至抗原及實質上相同的抗原，其意指具有10^-7M或更小、10^-8M或更小、5 x 10^-9M或更小、或介於10^-8M與10^-10M或更小之間之K_D，但不以高親和力結合至不相關的抗原。Antibodies typically bind specifically to their cognate antigen with high affinity, as reflected by a dissociation constant (_KD ) of^10-5 to^10-11 M or less. Any_KD greater than about^10-4 M is generally considered to indicate non-specific binding. As used herein, an antibody that "specifically binds" to an antigen refers to an antibody that binds to the antigen and substantially the same antigen with high affinity, meaning having a_KD of^10-7 M or less,^10-8 M or less, 5 x^10-9 M or less, or between^10-8 M and^10-10 M or less, but does not bind with high affinity to unrelated antigens.

免疫球蛋白可來自任何通常已知的同型物，包括(但不限於) IgA、分泌IgA、IgG及IgM。IgG同型物在某些物種中分為亞類：人類中之IgG1、IgG2、IgG3及IgG4、及小鼠中之IgG1、IgG2a、IgG2b及IgG3。在一些態樣中，本文描述之抗CD161抗體為IgG1亞型。免疫球蛋白(例如IgG1)以幾種同種異型存在，其至多在幾個胺基酸中彼此不同。「抗體」包括(以實例說明之)天然存在及非天然存在之抗體；單株及多株抗體；嵌合及人類化抗體；人類及非人類的抗體及全合成抗體。Immunoglobulins can be from any commonly known isotype, including (but not limited to) IgA, secretory IgA, IgG and IgM. IgG isotypes are divided into subclasses in certain species: IgG1, IgG2, IgG3 and IgG4 in humans, and IgG1, IgG2a, IgG2b and IgG3 in mice. In some aspects, the anti-CD161 antibodies described herein are of the IgG1 subtype. Immunoglobulins (e.g., IgG1) exist in several isotypes that differ from each other in at most a few amino acids. "Antibodies" include (by way of example) naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human and non-human antibodies and fully synthetic antibodies.

術語抗體之「抗原結合部分」如本文所用係指抗體之保留特異性結合至抗原(例如人類CD161)之能力之一或多個片段。已顯示抗體之抗原結合功能可藉由全長抗體之片段進行。涵蓋於術語抗體(例如本文描述之抗CD161抗體)之「抗原結合部分」內之結合片段之實例包括(i) Fab片段(來自木瓜蛋白酶裂解之片段)或由V_L、V_H、LC及CH1域組成之類似單價片段；(ii) F(ab')2片段(來自胃蛋白酶裂解之片段)或包含在鉸鏈區經雙硫橋連接之兩個Fab片段之類似二價片段；(iii)由V_H及CH1域組成之Fd片段；(iv)由抗體之單一臂之V_L及V_H域組成之Fv片段；(v) dAb片段(Ward等人，(1989)Nature341:544-546)，其由V_H域組成；(vi)分離的互補決定區(CDR)及(vii)可視情況經合成連接子接合之兩個或更多個分離的CDR之組合。此外，儘管Fv片段之兩個域V_L及V_H由單獨基因編碼，但其可使用重組方法經合成連接子接合，該合成連接子使得其製成為其中V_L及V_H區配對以形成單價分子之單蛋白鏈(稱為單鏈Fv (scFv)；參見，例如，Bird等人(1988)Science242:423-426；及Huston等人(1988)Proc. Natl. Acad. Sci. USA85:5879-5883)。此類單鏈抗體亦意欲涵蓋於術語抗體之「抗原結合部分」內。此等抗體片段使用熟習此項技術者已知的習知技術來獲得，且以與完整抗體相同的方式針對效用篩選該等片段。抗原結合部分可藉由重組DNA技術、或藉由完整免疫球蛋白之酶促或化學裂解來產生。The term "antigen-binding portion" of an antibody as used herein refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (eg, human CD161). It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" of an antibody (e.g., the anti-CD161 antibodies described herein) include (i) a Fab fragment (fragment derived from papain cleavage) or a similar monovalent fragment consisting of the_VL ,_VH , LC, and CH1 domains; (ii) a F(ab')2 fragment (fragment derived from pepsin cleavage) or a similar bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the_VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of the antibody; (v) a dAb fragment (Ward et al., (1989)Nature 341:544-546), which consists of the_VL and_VH domains of the antibody; (vi) a separate complementary determining region (_CDR ) and (vii) a combination of two or more separate CDRs optionally joined by a synthetic linker. In addition, although the two domains of the Fv fragment,_VL and_VH, are encoded by separate genes, they can be joined by a synthetic linker using recombinant methods, which allows them to be made into a single protein chain in which the_VL and_VH regions pair to form a monovalent molecule (called a single-chain Fv (scFv); see, e.g., Bird et al. (1988)Science 242:423-426; and Huston et al. (1988)Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and are screened for utility in the same manner as intact antibodies. Antigen-binding portions may be produced by recombinant DNA techniques, or by enzymatic or chemical cleavage of intact immunoglobulins.

「雙特異性抗體」或「雙功能抗體」係具有兩個不同重鏈/輕鏈對及兩個不同結合位點之人工雜合抗體。「多特異性抗體」或「多功能抗體」係具有多於兩個不同重鏈/輕鏈對及兩個不同結合位點之人工雜合抗體。雙特異性及多特異性抗體可藉由多種方法(包括雜交瘤之融合或Fab'片段之連接)產生。參見，例如，Songsivilai及Lachmann,Clin. Exp. Immunol. 79:315-321 (1990)；Kostelny等人，J. Immunol. 148, 1547-1553 (1992)。"Bispecific antibodies" or "bifunctional antibodies" are artificial hybrid antibodies with two different heavy chain/light chain pairs and two different binding sites. "Multispecific antibodies" or "multifunctional antibodies" are artificial hybrid antibodies with more than two different heavy chain/light chain pairs and two different binding sites. Bispecific and multispecific antibodies can be produced by a variety of methods, including fusion of hybridomas or connection of Fab' fragments. See, e.g., Songsivilai and Lachmann,Clin. Exp. Immunol . 79:315-321 (1990); Kostelny et al.,J. Immunol . 148, 1547-1553 (1992).

術語「單株抗體」如本文所用係指來自實質上同源之抗體群體之抗體，亦即，群體中包含的個別抗體係實質上相似且且結合該(等)相同抗原決定基(例如該等抗體展現單一結合特異性及親和力)，但在產生該單株抗體期間可能產生的可能的變異體除外，此類變異體一般以少量存在。修飾語「單株」指示抗體之特徵，其係自實質上同源抗體群體獲得，且不應被解釋為需要藉由任一特定方法來產生抗體。術語「人類單株抗體」係指來自展現單一結合特異性之實質上同源抗體群體之抗體，且其具有衍生自人類生殖系免疫球蛋白序列之可變及可選恆定區。在一些態樣中，由雜交瘤產生人類單株抗體，該雜交瘤包括自轉殖基因非人類動物(例如轉殖基因小鼠)獲得之B細胞，其具有包含融合至永生化細胞之人類重鏈轉殖基因及輕鏈轉殖基因之基因組。The term "monoclonal antibody" as used herein refers to an antibody from a population of substantially homogeneous antibodies, that is, the individual antibodies contained in the population are substantially similar and bind to the same antigenic determinant(s) (e.g., the antibodies exhibit a single binding specificity and affinity), except for possible variants that may arise during the production of the monoclonal antibody, such variants generally being present in minor amounts. The modifier "monoclonal" indicates the characteristic of the antibody that it is obtained from a population of substantially homogeneous antibodies and should not be construed as requiring the antibody to be produced by any particular method. The term "human monoclonal antibody" refers to an antibody from a population of substantially homogeneous antibodies that exhibit a single binding specificity and that has variable and selectable constant regions derived from human germline immunoglobulin sequences. In some aspects, human monoclonal antibodies are produced by a hybridoma comprising B cells obtained from a transgenic non-human animal (e.g., a transgenic mouse) having a genome comprising a human heavy chain transgene and a light chain transgene fused to an immortalized cell.

術語「重組人類抗體」如本文所用包括藉由重組手段製備、表現、創建或分離的所有人類抗體，諸如(a)自對於人類免疫球蛋白基因或自其製備的雜交瘤而言係轉殖基因或轉染色體之動物(例如小鼠)分離的抗體；(b)自經轉化以表現該抗體例如自轉染瘤(transfectoma)之宿主細胞分離的抗體；(c)自重組、組合人類抗體庫分離的抗體；及(d)藉由涉及將人類免疫球蛋白基因序列剪接至其他DNA序列之任何其他手段製備、表現、創建或分離的抗體。此類重組人類抗體包含利用由生殖系基因編碼之特定人類生殖系免疫球蛋白序列之可變區及恆定區，但包括例如在抗體成熟期間發生之序列後續重排及突變。如此項技術中已知(參見，例如，Lonberg (2005)Nature Biotech. 23(9): 1117-1125)，可變區含有抗原結合域，其由各種基因編碼，該等基因經重排以形成對外來抗原具有特異性之抗體。除了重排外，可變區可藉由多個單胺基酸變化(稱為體細胞突變或超突變)進一步修飾以增加該抗體對外來抗原之親和力。恆定區將進一步回應於抗原而改變(亦即同型物切換)。因此，回應於抗原而編碼輕鏈及重鏈免疫球蛋白多肽之經重排及體細胞突變之核酸分子不能與原始核酸分子具有序列一致性，而是將實質上相同或相似(亦即具有至少80%一致性)。The term "recombinant human antibody" as used herein includes all human antibodies prepared, expressed, created or isolated by recombinant means, such as (a) antibodies isolated from animals (e.g., mice) that are transgenic or transchromosomal for human immunoglobulin genes or from hybridomas prepared therefrom; (b) antibodies isolated from host cells transformed to express the antibody, such as from transfectomas; (c) antibodies isolated from recombinant, combinatorial human antibody libraries; and (d) antibodies prepared, expressed, created or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies comprise variable and constant regions that utilize specific human germline immunoglobulin sequences encoded by germline genes, but include subsequent rearrangement and mutation of sequences that occur, for example, during antibody maturation. As is known in the art (see, e.g., Lonberg (2005)Nature Biotech . 23(9): 1117-1125), the variable region contains an antigen binding domain that is encoded by various genes that are rearranged to form antibodies specific for foreign antigens. In addition to rearrangement, the variable region can be further modified by multiple single amino acid changes (referred to as somatic mutations or hypermutations) to increase the affinity of the antibody for foreign antigens. The constant region will further change in response to the antigen (i.e., isotype switching). Thus, rearranged and somatically mutated nucleic acid molecules encoding light and heavy chain immunoglobulin polypeptides in response to an antigen must not have sequence identity to the original nucleic acid molecule, but will be substantially identical or similar (i.e., have at least 80% identity).

「人類」抗體(HuMAb)係指具有其中框架及CDR區衍生自人類生殖系免疫球蛋白序列之可變區之抗體。此外，若抗體含有恆定區，則該恆定區亦衍生自人類生殖系免疫球蛋白序列。本文描述之抗CD161抗體可包括非人類生殖系免疫球蛋白序列編碼之胺基酸殘基(例如藉由活體外隨機或定點誘變或藉由活體內體細胞突變引入之突變)。然而，術語「人類抗體」如本文所用意欲包括其中衍生自另一哺乳動物物種(諸如小鼠)之生殖系之CDR序列已接枝至人類框架序列上之抗體。該等術語「人類」抗體及「完全人類」抗體係同義使用。"Human" antibody (HuMAb) refers to an antibody having a variable region in which the framework and CDR regions are derived from human germline immunoglobulin sequences. In addition, if the antibody contains a constant region, the constant region is also derived from a human germline immunoglobulin sequence. The anti-CD161 antibody described herein may include amino acid residues encoded by non-human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by in vivo endosomal cell mutagenesis). However, the term "human antibody" as used herein is intended to include antibodies in which the CDR sequences derived from the germline of another mammalian species (such as mice) have been grafted onto human framework sequences. The terms "human" antibody and "fully human" antibody are used synonymously.

「人類化」抗體係指其中非人類的抗體之CDR域外部的一些、大部分或全部胺基酸經衍生自人類免疫球蛋白之對應胺基酸取代之抗體。在抗體之人類化形式之一些態樣中，CDR域外部的一些、大部分或全部胺基酸已經來自人類免疫球蛋白之胺基酸取代，而一或多個CDR區內的一些、大部分或全部胺基酸未改變。胺基酸之小量添加、缺失、插入、取代或修飾係允許的，只要其不廢除該抗體結合至特定抗原之能力即可。「人類化」抗體保留類似於原始抗體之抗原特異性之抗原特異性。A "humanized" antibody is one in which some, most, or all of the amino acids outside the CDR domains of a non-human antibody are replaced with corresponding amino acids derived from human immunoglobulins. In some aspects of humanized forms of antibodies, some, most, or all of the amino acids outside the CDR domains have been replaced with amino acids from human immunoglobulins, while some, most, or all of the amino acids within one or more CDR regions are not altered. Minor additions, deletions, insertions, substitutions, or modifications of amino acids are permitted as long as they do not abolish the ability of the antibody to bind to a specific antigen. A "humanized" antibody retains an antigenic specificity similar to that of the original antibody.

「嵌合抗體」係指其中可變區衍生自一個物種且恆定區衍生自另一物種之抗體，諸如其中該等可變區衍生自小鼠抗體且該等恆定區衍生自人類抗體之抗體。"Chimeric antibodies" refer to antibodies in which the variable regions are derived from one species and the constant regions are derived from another species, such as antibodies in which the variable regions are derived from a mouse antibody and the constant regions are derived from a human antibody.

如本文所用，「同型物」係指由重鏈恆定區基因編碼之抗體類別(例如IgG1、IgG2、IgG3、IgG4、IgM、IgA1、IgA2、IgD及IgE抗體)。As used herein, "isotype" refers to the antibody class encoded by the heavy chain constant region gene (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE antibodies).

片語「識別抗原之抗體」及「對抗原具有特異性之抗體」在本文中可與術語「特異性結合至抗原之抗體」互換使用。The phrases "antibodies that recognize antigens" and "antibodies specific for antigens" are used interchangeably herein with the term "antibodies that specifically bind to antigens."

「分離的抗體」如本文所用意欲指實質上不含其他蛋白質及細胞材料之抗體。"Isolated antibody" as used herein is intended to refer to an antibody that is substantially free of other proteins and cellular materials.

如本文所用，術語「CD161」係指由人類KLRB1基因編碼之多肽，其在NK細胞子集且在周邊血液T細胞中表現。CD161亦可稱為「殺手細胞凝集素樣受體亞家族B成員1」、「KLRB1」、「C型凝集素域家族5成員B」、「CLEC5B」、「C型凝集素域家族5成員B」及「NKRP1a」。CD161被認為藉由結合CLEC2D/LLT1而抑制靶細胞中之NK細胞介導之細胞毒性及干擾素-γ分泌。經活化之CD161導致特異性酸性神經髓磷脂酶(aSMase)刺激及細胞內神經醯胺之後續顯著升高。CD161充當結合至末端碳水化合物Gal-α (1,3)Gal抗原決定基以及結合至N-乙醯半乳糖胺抗原決定基之凝集素。典型人類CD161序列提供於表1 (UniProt Q12918)中。As used herein, the term "CD161" refers to a polypeptide encoded by the human KLRB1 gene, which is expressed in NK cell subsets and in peripheral blood T cells. CD161 may also be referred to as "killer cell lectin-like receptor subfamily B member 1", "KLRB1", "C-type lectin domain family 5 member B", "CLEC5B", "C-type lectin domain family 5 member B", and "NKRP1a". CD161 is believed to inhibit NK cell-mediated cytotoxicity and interferon-γ secretion in target cells by binding to CLEC2D/LLT1. Activated CD161 leads to specific acidic nerve myelinase (aSMase) stimulation and subsequent significant increase in intracellular neuramide. CD161 acts as a lectin that binds to the terminal carbohydrate Gal-α (1,3)Gal epitope and to the N-acetylgalactosamine epitope. A typical human CD161 sequence is provided in Table 1 (UniProt Q12918).

表1：典型人類CD161胺基酸序列。Table1: Typical human CD161 amino acid sequence.

如本文所用，抑制「CD161活性」之抗體意欲指抑制或減少CD161之一或多種活性之抗體。在一些態樣中，本文揭示之抗CD161抗體抑制或減少人類CD161與CLEC2D之間的相互作用。在一些態樣中，本文揭示之抗CD161抗體增加NK細胞介導之細胞毒性，亦即，藉由移除CD161介導之抑制。在一些態樣中，本文揭示之抗CD161抗體增加靶細胞干擾素-γ分泌，亦即，藉由移除CD161介導之抑制。在一些態樣中，本文揭示之抗CD161抗體抑制或減少靶細胞中之特異性酸性神經髓磷脂酶(aSMase)刺激。As used herein, an antibody that inhibits "CD161 activity" is intended to refer to an antibody that inhibits or reduces one or more activities of CD161. In some aspects, the anti-CD161 antibodies disclosed herein inhibit or reduce the interaction between human CD161 and CLEC2D. In some aspects, the anti-CD161 antibodies disclosed herein increase NK cell-mediated cytotoxicity, that is, by removing CD161-mediated inhibition. In some aspects, the anti-CD161 antibodies disclosed herein increase target cell interferon-γ secretion, that is, by removing CD161-mediated inhibition. In some aspects, the anti-CD161 antibodies disclosed herein inhibit or reduce specific acidic myelinase (aSMase) stimulation in target cells.

「Fc區」 (片段可結晶區)或「Fc域」或「Fc」係指C端區介導免疫球蛋白與宿主組織或因子(包括與位於免疫系統之各種細胞(例如效應細胞)上之Fc受體或至典型補體系統之第一組分(C1q))之結合之抗體之重鏈。因此，Fc區包含第一恆定區免疫球蛋白域(例如CH1或CL)除外的抗體之恆定區。在IgG、IgA及IgD抗體同型物中，Fc區包含衍生自該抗體的兩個重鏈之第二(CH2)及第三(CH3)恆定域之兩個相同蛋白質片段；IgM及IgE Fc區在每個多肽鏈中包含三個重鏈恆定域(CH域2至4)。對於IgG，Fc區包含免疫球蛋白域CH2及CH3及介於CH1域與CH2域之間的鉸鏈。儘管免疫球蛋白重鏈之Fc區之邊界的定義可能不同，如本文所定義，但人類IgG重鏈Fc區經定義成自IgG1的胺基酸殘基D221、IgG2的V222、IgG3的L221及IgG4的P224延伸至重鏈的羧基端，其中編碼係根據如在Kabat中之EU索引。人類IgG Fc區之CH2域自胺基酸237延伸至胺基酸340，且CH3域位於Fc區中之CH2域的C端側，亦即，其自胺基酸341延伸至IgG之胺基酸447或446 (若該C端離胺酸殘基不存在)或445 (若該C端甘胺酸及離胺酸殘基不存在)。如本文所用，Fc區可係天然序列Fc，包括任何同種異體變異體或變異體Fc (例如非天然存在之Fc)。"Fc region" (fragment crystallizable region) or "Fc domain" or "Fc" refers to the C-terminal region of an antibody's heavy chain that mediates the binding of the immunoglobulin to host tissues or factors, including to Fc receptors located on various cells of the immune system (e.g., effector cells) or to the first component (C1q) of the classical complement system. Thus, the Fc region comprises the constant region of an antibody excluding the first constant region immunoglobulin domain (e.g., CH1 or CL). In IgG, IgA and IgD antibody isotypes, the Fc region comprises two identical protein fragments derived from the second (CH2) and third (CH3) constant region domains of the two heavy chains of the antibody; IgM and IgE Fc regions comprise three heavy chain constant region domains (CH domains 2 to 4) in each polypeptide chain. For IgG, the Fc region comprises the immunoglobulin domains CH2 and CH3 and a hinge between the CH1 and CH2 domains. Although the definition of the boundaries of the Fc region of an immunoglobulin heavy chain may vary, as defined herein, the human IgG heavy chain Fc region is defined to extend from amino acid residues D221 of IgG1, V222 of IgG2, L221 of IgG3, and P224 of IgG4 to the carboxyl terminus of the heavy chain, wherein the numbering is according to the EU index as in Kabat. The CH2 domain of the human IgG Fc region extends from amino acid 237 to amino acid 340, and the CH3 domain is located on the C-terminal side of the CH2 domain in the Fc region, that is, it extends from amino acid 341 to amino acid 447 or 446 (if the C-terminal lysine residue is absent) or 445 (if the C-terminal glycine and lysine residues are absent) of IgG. As used herein, the Fc region may be a native sequence Fc, including any allotype variant or variant Fc (e.g., non-naturally occurring Fc).

「天然序列Fc區」或「天然序列Fc」包含與在自然界中發現的Fc區之胺基酸序列相同的胺基酸序列。天然序列人類Fc區包括天然序列人類IgG1 Fc區；天然序列人類IgG2 Fc區；天然序列人類IgG3 Fc區；及天然序列人類IgG4 Fc區以及其天然存在之變異體。天然序列Fc包括Fc之各種同種異型(參見，例如，Jefferis等人(2009)mAbs1: 1)。A "native sequence Fc region" or "native sequence Fc" comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. Native sequence human Fc regions include a native sequence human IgG1 Fc region; a native sequence human IgG2 Fc region; a native sequence human IgG3 Fc region; and a native sequence human IgG4 Fc region, and naturally occurring variants thereof. Native sequence Fc includes various isotypes of Fc (see, e.g., Jefferis et al. (2009)mAbs 1: 1).

術語「抗原決定基」或「抗原決定子」係指抗原(例如CD161)上的免疫球蛋白或抗體特異性結合的位點，例如，如用於識別其之特定方法所定義。抗原決定基可由鄰接胺基酸(通常係線性抗原決定基)或藉由蛋白質之三級折疊並列的非鄰接胺基酸(通常係構象抗原決定基)形成。由鄰接胺基酸形成的抗原決定基通常(但不總是)在暴露於變性溶劑時保留，而由三級折疊形成的抗原決定基通常在用變性溶劑處理時丟失。抗原決定基通常包括呈獨特空間構象的至少3、4、5、6、7、8、9、10、11、12、13、14或15個胺基酸。用於確定哪些抗原決定基藉由給定抗體鍵結(亦即抗原決定基定位)之方法係此項技術中熟知的且包括(例如)免疫轉漬及免疫沉澱分析，其中測試來自(例如來自CD161)之重疊或鄰接肽之與給定抗體(例如抗CD161抗體)之反應性。確定抗原決定基之空間構象的方法包括此項技術中之技術及彼等描述於其中者，例如x射線晶體學、x射線共晶體學、抗原突變分析、2維核磁共振及HDX-MS (參見，例如，Epitope Mapping Protocols in Methods in Molecular Biology, 第66卷, G. E. Morris編(1996))。The term "antigenic determinant" or "antigenic determinant" refers to a site on an antigen (e.g., CD161) to which an immunoglobulin or antibody specifically binds, for example, as defined by a particular method for identifying it. An antigenic determinant may be formed by adjacent amino acids (usually linear antigenic determinants) or by non-adjacent amino acids juxtaposed by tertiary folding of a protein (usually conformational antigenic determinants). Antigenic determinants formed by adjacent amino acids are usually (but not always) retained upon exposure to a denaturing solvent, while antigenic determinants formed by tertiary folding are usually lost upon treatment with a denaturing solvent. An antigenic determinant typically includes at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in a unique spatial conformation. Methods for determining which antigenic determinants are bound by a given antibody (i.e., antigenic determinant localization) are well known in the art and include, for example, immunoblotting and immunoprecipitation assays, in which overlapping or adjacent peptides from (e.g., from CD161) are tested for reactivity with a given antibody (e.g., anti-CD161 antibody). Methods for determining the spatial conformation of an antigenic determinant include techniques in the art and those described therein, such as x-ray crystallography, x-ray co-crystals, antigenic mutation analysis, 2-dimensional nuclear magnetic resonance, and HDX-MS (see, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, G. E. Morris, ed. (1996)).

術語「抗原決定基定位」係指識別抗體-抗原識別之分子決定基之製程。The term "antigenic determinant mapping" refers to the process of identifying the molecular determinants of antibody-antigen recognition.

提及兩個或更多個抗體之術語「結合至相同抗原決定基」意指該等抗體結合至胺基酸殘基之相同區段，藉由給定方法確定。用於確定抗體是否與本文描述之抗體結合至「CD161上的相同抗原決定基」之技術包括(例如)抗原決定基定位方法，諸如提供抗原決定基之原子解析度之抗原:抗體複合物之晶體之x射線分析及氫/氘交換質譜(HDX-MS)。其他方法監測該抗體對抗原片段或該抗原之突變變異體之結合，其中由於抗原序列內的胺基酸殘基之修飾所致之結合之損失經常被認為是抗原決定基組分之指示。此外，亦可使用用於抗原決定基定位之計算組合方法。此等方法依賴於所關注抗體自組合噬菌體展示肽庫親和力分離特異性短肽之能力。具有相同VH及VL或相同CDR1、2及3序列之抗體預期會結合至相同抗原決定基。The term "binding to the same antigenic determinant" referring to two or more antibodies means that the antibodies bind to the same segment of amino acid residues, as determined by a given method. Techniques for determining whether an antibody binds to the "same antigenic determinant on CD161" as the antibodies described herein include, for example, antigenic determinant localization methods, such as x-ray analysis and hydrogen/deuterium exchange mass spectrometry (HDX-MS) of crystals of antigen:antibody complexes that provide atomic resolution of the antigenic determinant. Other methods monitor the binding of the antibody to antigenic fragments or mutant variants of the antigen, where loss of binding due to modification of amino acid residues within the antigenic sequence is often considered to be an indication of antigenic determinant composition. In addition, computational combinatorial methods for antigenic determinant localization can also be used. These methods rely on the ability of the antibody of interest to affinity isolate specific short peptides from an assemblage of phage-displayed peptide libraries. Antibodies with identical VH and VL or identical CDR1, 2, and 3 sequences are expected to bind to the same antigenic determinant.

「與另一抗體競爭結合至靶」之抗體係指抑制(部分或完全)另一抗體至該靶之結合之抗體。可使用已知競爭實驗(例如BIACORE^®表面電漿子共振(SPR)分析)來確定兩種抗體是否彼此競爭結合至靶，亦即，一種抗體是否及在何種程度上抑制另一抗體至靶之結合。在一些態樣中，抗體與另一抗體競爭且抑制另一抗體至靶之結合至少50%、60%、70%、80%、90%或100%。抑制或競爭之程度可取決於哪種抗體係「阻斷抗體」 (亦即首先與靶一起培養之冷抗體)而有所不同。競爭分析可如例如Ed Harlow及David Lane, Cold Spring Harb Protoc; 2006；doi: 10.1101/pdb.prot4277、或Ed Harlow及David Lane之章節11的「Using Antibodies」, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999中所述進行。若抗體彼此雙向阻斷至少50%，亦即無論一種或另一抗體是否在該競爭實驗中首先與該抗原接觸，則兩種抗體「交叉競爭」。An antibody that "competes with another antibody for binding to a target" refers to an antibody that inhibits (partially or completely) the binding of another antibody to the target. Known competition assays (e.g.,^BIACORE® surface plasmon resonance (SPR) analysis) can be used to determine whether two antibodies compete with each other for binding to a target, that is, whether and to what extent one antibody inhibits the binding of the other antibody to the target. In some aspects, an antibody competes with the other antibody and inhibits the binding of the other antibody to the target by at least 50%, 60%, 70%, 80%, 90%, or 100%. The degree of inhibition or competition may vary depending on which antibody is the "blocking antibody" (i.e., the cold antibody that is first incubated with the target). Competition assays can be performed as described, for example, in Ed Harlow and David Lane, Cold Spring Harb Protoc; 2006; doi: 10.1101/pdb.prot4277, or in Chapter 11 of Ed Harlow and David Lane, "Using Antibodies", Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999. Two antibodies "cross-compete" if they block each other bidirectionally by at least 50%, i.e., regardless of whether one or the other antibody is first exposed to the antigen in the competition experiment.

用於確定兩種抗體是否競爭或交叉競爭結合之競爭性結合分析包括：競爭結合至表現CD161之細胞，例如藉由流動式細胞測量術，諸如實例中所描述。其他方法包括：SPR (例如BIACORE^®)、固相直接或間接放射免疫分析(RIA)、固相直接或間接酵酵素免疫分析(EIA)、夾心競爭分析(參見Stahli等人，Methods in Enzymology9:242 (1983))；固相直接生物素-抗生物素蛋白EIA (參見Kirkland等人，J. Immunol. 137:3614 (1986))；固相直接標記分析、固相直接標記夾心分析(參見Harlow及Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988))；使用1-125標記的固相直接標記RIA (參見Morel等人，Mol. Immunol. 25(1):7 (1988))；固相直接生物素-抗生物素蛋白EIA (Cheung等人，Virology176:546 (1990))；及直接標記RIA.(Moldenhauer等人，Scand. J. Immunol. 32:77 (1990))。Competitive binding assays used to determine whether two antibodies compete or cross-compete for binding include competitive binding to cells expressing CD161, such as by flow cytometry, as described in the Examples. Other methods include: SPR (e.g.,^BIACORE® ), solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see Stahli et al.,Methods in Enzymology 9:242 (1983)); solid phase direct biotin-avidin EIA (see Kirkland et al.,J. Immunol . 137:3614 (1986)); solid phase direct label assay, solid phase direct label sandwich assay (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)); solid phase direct label RIA using 1-125 label (see Morel et al.,Mol. Immunol . 25(1):7 (1988)); solid phase direct biotin-avidin EIA (Cheung et al.,Virology 176:546 (1990)); and direct labeling RIA. (Moldenhauer et al.,Scand. J. Immunol . 32:77 (1990)).

如本文所用，術語「特異性結合(specific binding)」、「選擇性結合(selective binding)」、「選擇性結合(selectively binds)」及「特異性結合(specifically binds)」係指結合至預先確定之抗原上的抗原決定基之抗體。通常，該抗體(i)以約小於10^-7M，諸如約小於10^-8M、10^-9M或10^-10M或甚至更低之平衡解離常數(K_D)結合，當藉由例如表面電漿子共振(SPR)技術在BIACORE^®2000儀器中使用該預先確定之抗原(例如重組人類CD161)作為該分析物及使用該抗體作為配位體、或該抗體對抗原陽性細胞之結合之Scatchard分析測定時，且(ii)以至少兩倍大於其結合至非特異性抗原(例如BSA、酪蛋白)而非預先確定之抗原或緊密相關抗原之親和力之親和力結合至預先確定之抗原。因此，「特異性結合至CD161」之抗體係指以10^-7M或更小，諸如約小於10^-8M、10^-9M或10^-10M或甚至更低之K_D結合至CD161之抗體。在一些態樣中，在標準結合分析中，此類不與來自非人類的物種之CD161交叉反應之抗體展現對此等蛋白質之基本上不可偵測之結合。As used herein, the terms "specific binding,""selectivebinding,""selectivelybinds," and "specifically binds" refer to antibodies that bind to an antigenic determinant on a predetermined antigen. Typically, the antibody (i) binds with an equilibrium dissociation constant (_KD ) of less than about^10-7 M, such as less than about^10-8 M,^10-9 M or^10-10 M or even lower, when measured by, for example, surface plasmon resonance (SPR) technology in a^BIACORE® 2000 instrument using the predetermined antigen (e.g., recombinant human CD161) as the analyte and the antibody as a ligand, or Scatchard analysis of binding of the antibody to antigen-positive cells, and (ii) binds to the predetermined antigen with an affinity that is at least two-fold greater than its affinity for binding to a non-specific antigen (e.g., BSA, casein) other than the predetermined antigen or a closely related antigen. Thus, an antibody that "specifically binds to CD161" refers to an antibody that binds to CD161 with a_KD of^10-7 M or less, such as less than about^10-8 M,^10-9 M, or^10-10 M, or even less. In some aspects, such antibodies that do not cross-react with CD161 from non-human species exhibit essentially undetectable binding to these proteins in standard binding assays.

術語「k_assoc」或「k_a」如本文所用意欲指特定抗體-抗原相互作用之締合速率，而術語「k_dis」或「k_d」如本文所用意欲指特定抗體-抗原相互作用之解離速率。術語「K_D」如本文所用意欲指解離常數，其自k_d與k_a之比(亦即k_d/k_a)獲得且表示為莫耳濃度(M)。可使用此項技術中明確確立的方法來測定抗體之K_D值。用於確定抗體之K_D之可用方法包括表面電漿子共振、生物感測系統(諸如BIACORE^®系統)或流動式細胞測量術及Scatchard分析。The term "_kassoc " or "_ka " as used herein is intended to refer to the association rate of a particular antibody-antigen interaction, while the term "_kdis " or "_kd " as used herein is intended to refer to the dissociation rate of a particular antibody-antigen interaction. The term "_KD " as used herein is intended to refer to the dissociation constant, which is obtained from the ratio of_kd to_ka (i.e.,_kd /_ka ) and expressed as molar concentration (M). The_KD value of an antibody can be determined using well-established methods in the art. Available methods for determining the_KD of an antibody include surface plasmon resonance, biosensing systems (such as the^BIACORE® system) or flow cytometry and Scatchard analysis.

如本文所用，術語IgG抗體之「高親和力」係指對靶抗原具有10^-8M或更小、10^-9M或更小、或10^-10M或更小之K_D之抗體。然而，「高親和力」結合因其他抗體同型物而異。例如，IgM同型物之「高親和力」結合係指具有10^-10M或更小、或10^-8M或更小之K_D之抗體。As used herein, the term "high affinity" for an IgG antibody refers to an antibody that has a_KD of^10-8 M or less,^10-9 M or less, or^10-10 M or less for the target antigen. However, "high affinity" binding varies for other antibody isotypes. For example, "high affinity" binding for an IgM isotype refers to an antibody that has a_KD of^10-10 M or less, or^10-8 M or less.

術語「EC₅₀」在使用抗體或其抗原結合片段之活體外或活體內分析之上下文中係指誘導為最大反應的50%的反應之抗體或其抗原結合部分之濃度，亦即介於最大反應與基線之間的半路。The term "_EC50 " in the context of an in vitro or in vivo assay using an antibody or antigen-binding fragment thereof refers to the concentration of the antibody or antigen-binding portion thereof that induces a response that is 50% of the maximal response, ie, halfway between the maximal response and baseline.

術語「天然存在之」如本文中應用於物體時所用係指物體可在自然界中發現之事實。例如，存在於生物(包括病毒)中之可自天然來源分離且尚未經人類在實驗室中故意修飾之多肽或多核苷酸序列係天然存在的。The term "naturally occurring" as used herein when applied to an object refers to the fact that an object can be found in nature. For example, a polypeptide or polynucleotide sequence present in an organism (including a virus) that can be isolated from a natural source and has not been intentionally modified by humans in the laboratory is naturally occurring.

「多肽」係指包含至少兩個連續連接之胺基酸殘基之鏈，其中於該鏈之長度上沒有上限。蛋白質中之一或多個胺基酸殘基可含有修飾，諸如(但不限於)醣基化、磷酸化或雙硫鍵形成。「蛋白質」可包含一或多種多肽。A "polypeptide" refers to a chain of at least two consecutively linked amino acid residues, wherein there is no upper limit on the length of the chain. One or more of the amino acid residues in a protein may contain modifications, such as (but not limited to) glycosylation, phosphorylation or disulfide bond formation. A "protein" may include one or more polypeptides.

術語「核酸分子」如本文所用意欲包括DNA分子及RNA分子。核酸分子可係單股或雙股，且可係cDNA。The term "nucleic acid molecule" as used herein is intended to include DNA molecules and RNA molecules. The nucleic acid molecule may be single-stranded or double-stranded, and may be cDNA.

「保守胺基酸取代」係指胺基酸殘基經具有相似側鏈之胺基酸殘基之取代。具有相似側鏈之胺基酸殘基之家族已在此項技術中定義。此等家族包括具有鹼性側鏈 (例如離胺酸、精胺酸、組胺酸)、酸性側鏈(例如天冬胺酸、麩胺酸)、不帶電荷之極性側鏈(例如甘胺酸、天冬醯胺酸、麩醯胺酸、絲胺酸、蘇胺酸、酪胺酸、半胱胺酸、色胺酸)、非極性側鏈(例如丙胺酸、纈胺酸、白胺酸、異白胺酸、脯胺酸、苯丙胺酸、甲硫胺酸)、β-分支鏈側鏈(例如蘇胺酸、纈胺酸、異白胺酸)及芳族側鏈(例如酪胺酸、苯丙胺酸、色胺酸、組胺酸)之胺基酸。在一些態樣中，將抗CD161抗體中之預測非必需胺基酸殘基改由來自相同側鏈家族之另一胺基酸殘基置換。識別不消除抗原結合之核苷酸及胺基酸保守取代之方法係此項技術中熟知的(參見，例如，Brummell等人，Biochem. 32: 1180-1187 (1993)；Kobayashi等人，Protein Eng. 12(10):879-884 (1999)；及Burks等人，Proc. Natl. Acad. Sci. USA94:412-417 (1997))。"Conservative amino acid substitution" refers to the replacement of an amino acid residue with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamine), uncharged polar side chains (e.g., glycine, aspartic acid, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). In some aspects, the predicted non-essential amino acid residue in the anti-CD161 antibody is replaced by another amino acid residue from the same side chain family. Methods for identifying nucleotides and amino acid conservative substitutions that do not eliminate antigen binding are well known in the art (see, e.g., Brummell et al.,Biochem . 32: 1180-1187 (1993); Kobayashi et al.,Protein Eng . 12(10): 879-884 (1999); and Burks et al.,Proc. Natl. Acad. Sci. USA 94: 412-417 (1997)).

對於核酸，術語「實質同源性」指示兩個核酸或其指定序列在至少約80%之核苷酸、至少約90%至95%、或至少約98%至99.5%之核苷酸中在最佳比對及比較時係相同的，具有適宜核苷酸插入或缺失。或者，當區段將在選擇性雜交條件下雜交至股之補體時存在實質同源性。For nucleic acids, the term "substantial homology" indicates that two nucleic acids or designated sequences thereof are identical in at least about 80% of the nucleotides, at least about 90% to 95%, or at least about 98% to 99.5% of the nucleotides when optimally aligned and compared, with appropriate nucleotide insertions or deletions. Alternatively, substantial homology exists when the segments will hybridize to the complement of a strand under selective hybridization conditions.

對於多肽，術語「實質同源性」指示兩個多肽或其指定序列在至少約80%之胺基酸、至少約90%至95%、或至少約98%至99.5%之胺基酸中在最佳比對及比較時係相同的，具有適宜胺基酸插入或缺失。With respect to polypeptides, the term "substantial homology" indicates that two polypeptides or designated sequences thereof are identical in at least about 80% of the amino acids, at least about 90% to 95%, or at least about 98% to 99.5% of the amino acids when optimally aligned and compared, with appropriate amino acid insertions or deletions.

兩個序列間之一致性百分比係考慮到空位數及每個空位之長度(需要引入其用於兩個序列之最佳比對)，該等序列共有的相同位置數目(亦即%同源性 = 相同位置# /總位置# x 100)的函數。兩個序列間之序列之比較及一致性百分比之測定可使用數學演算法來達成，如下文在非限制性實例中所述。The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., % homology = # of identical positions / # of total positions x 100), taking into account the number of gaps and the length of each gap (which needs to be introduced for optimal alignment of the two sequences). The comparison of sequences and determination of percent identity between two sequences can be achieved using a mathematical algorithm, as described below in the non-limiting examples.

兩個核苷酸序列間之一致性百分比可使用GCG軟體包(可在worldwideweb.gcg.com下取得)中之GAP程式，使用NWSgapdna.CMP矩陣及40、50、60、70或80之空位權重及1、2、3、4、5或6之長度權重來測定。兩個核苷酸或胺基酸序列間之一致性百分比亦可使用E. Meyers及W. Miller (CABIOS, 4: 11-17 (1989))之演算法來測定，該演算法已使用PAM120權數殘基表、12之空位長度罰分及4之空位罰分併入至ALIGN程式(版本2.0)中。此外，兩個胺基酸序列間之一致性百分比可使用Needleman及Wunsch (J. Mol.Biol. (48):444-453 (1970))演算法來測定，該演算法已使用Blossum 62矩陣或PAM250矩陣、及16、14、12、10、8、6或4之空位權重及1、2、3、4、5或6之長度權重併入至GCG軟體包(可在http://www.gcg.com下取得)之GAP程式中。The percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at worldwideweb.gcg.com) using the NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. The percent identity between two nucleotide or amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (CABIOS , 4: 11-17 (1989)), which has been incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. In addition, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol . (48): 444-453 (1970)) algorithm, which has been incorporated into the GAP program in the GCG software package (available at http://www.gcg.com) using either the Blossum 62 matrix or the PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.

本文所述之核酸及蛋白質序列可進一步用作「查詢序列」來進行針對公開數據庫的搜索以例如識別相關序列。此類搜索可使用Altschul等人(1990)J. Mol. Biol. 215:403-10之NBLAST及XBLAST程式(版本2.0)來進行。BLAST核苷酸搜索可利用NBLAST程式，分數 = 100，字長 = 12進行以達成與本文所述之核酸分子同源之核苷酸序列。BLAST蛋白質搜索可利用XBLAST程式，分數 = 50，字長 = 3進行以達成與本文所述之蛋白質分子同源之胺基酸序列。為了達成用於比較目的之空位比對，帶空位(Gapped) BLAST可如Altschul等人，(1997)Nucleic Acids Res. 25(17):3389-3402中所述進行使用。當使用BLAST及帶空位BLAST程式時，可使用該相應程式(例如XBLAST及NBLAST)之預設參數。參見worldwideweb.ncbi.nlm.nih.gov。The nucleic acid and protein sequences described herein can further be used as "query sequences" to perform searches against public databases, e.g., to identify related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul et al. (1990)J. Mol. Biol . 215:403-10. BLAST nucleotide searches can be performed using the NBLAST program, score = 100, word length = 12 to achieve nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed using the XBLAST program, score = 50, word length = 3 to achieve amino acid sequences homologous to the protein molecules described herein. To achieve gapped alignments for comparison purposes, Gapped BLAST can be used as described in Altschul et al., (1997)Nucleic Acids Res . 25(17):3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the corresponding programs (e.g., XBLAST and NBLAST) can be used. See worldwideweb.ncbi.nlm.nih.gov.

核酸可存在於整個細胞、細胞溶解產物中、或以部分純化或實質上純淨之形式存在。核酸在藉由標準技術(包括鹼性/SDS處理、CsCl顯帶(CsCl banding)、管柱層析、瓊脂糖凝膠電泳及此項技術中熟知的其他技術)自其他細胞組分或其他污染物(例如其他細胞核酸(例如該染色體之其他部分)或蛋白質)純化時「經分離」或「致使變得實質上純淨」。參見，F. Ausubel等人編, Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987)。Nucleic acids may be present in whole cells, in cell lysates, or in partially purified or substantially pure form. Nucleic acids are "isolated" or "rendered substantially pure" when they are purified from other cellular components or other contaminants, such as other cellular nucleic acids (e.g., other portions of the chromosome) or proteins, by standard techniques, including alkaline/SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other techniques well known in the art. See, F. Ausubel et al., eds., Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987).

核酸(例如cDNA)可根據標準技術進行突變以提供基因序列。對於編碼序列，此等突變可根據需要影響胺基酸序列。特定言之，設想與天然V、D、J、常數、切換及本文描述之其他此類序列實質上同源或衍生自天然V、D、J、常數、切換及本文描述之其他此類序列之DNA序列(其中「衍生」指示與另一序列相同或自另一序列修飾之序列)。Nucleic acids (e.g., cDNA) can be mutated according to standard techniques to provide gene sequences. For coding sequences, such mutations can affect the amino acid sequence as desired. In particular, DNA sequences substantially homologous to or derived from natural V, D, J, constant, switch, and other such sequences described herein are contemplated (where "derived" indicates a sequence that is identical to or modified from another sequence).

術語「載體」如本文所用意欲指能夠運輸其已連接的另一核酸之核酸分子。載體之一種類型係「質體」，其係指其中可接合另外DNA區段之環狀雙股DNA環。載體之另一種類型係病毒載體，其中可將另外DNA區段接合至該病毒基因組中。某些載體能夠在其所引入的宿主細胞中自主複製(例如具有細菌複製起點之細菌載體、及游離型哺乳動物載體)。在引入至宿主細胞中時，可將其他載體(例如非游離型哺乳動物載體)整合至宿主細胞之該基因組中，且由此連同該宿主基因組一起進行複製。此外，某些載體能夠引導其所操作連接的基因之表現。此類載體在本文中稱為「重組表現載體」(或簡稱「表現載體」)。一般而言，重組DNA技術中之效用之表現載體經常呈質體之形式。在本說明書中，「質體」及「載體」可互換使用，因為該質體係載體之最常用的形式。然而，亦包括表現載體之其他形式，諸如病毒載體(例如複製缺陷逆轉錄病毒、腺病毒及腺相關病毒)，其提供等效功能。The term "vector" as used herein is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plastid," which refers to a circular double-stranded DNA loop into which additional DNA segments can be joined. Another type of vector is a viral vector, into which additional DNA segments can be joined to the viral genome. Certain vectors are capable of autonomous replication in the host cell into which they are introduced (e.g., bacterial vectors with a bacterial replication origin, and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of the host cell upon introduction into the host cell, and thereby replicated along with the host genome. In addition, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). In general, expression vectors useful in recombinant DNA technology are often in the form of plasmids. In this specification, "plasmid" and "vector" are used interchangeably, as the plasmid is the most commonly used form of vector. However, other forms of expression vectors, such as viral vectors (e.g., replication-defective retroviruses, adenoviruses, and adeno-associated viruses), are also included, which provide equivalent functions.

術語「重組宿主細胞」(或簡稱為「宿主細胞」)如本文所用意欲指包含非天然存在於該細胞中之核酸之細胞，且可係已引入重組表現載體的細胞。應理解，此類術語無意僅指特定個體細胞而且指此一細胞之子代。因為某些修飾可由於變或環境影響而在後代中發生，此種後代可能事實上與親本細胞不同，但仍包括在如本文所用的術語「宿主細胞」之範疇內。The term "recombinant host cell" (or simply "host cell") as used herein is intended to refer to a cell that contains a nucleic acid that is not naturally present in the cell, and may be a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are not intended to refer only to a particular individual cell but also to the progeny of such a cell. Because certain modifications may occur in subsequent generations due to mutation or environmental influences, such progeny may not actually be the same as the parent cell, but are still included in the scope of the term "host cell" as used herein.

如本文所用，「投與」係指使用熟習此項技術者已知的各種方法及遞送系統中之任何者，將包含治療劑之組合物物理引入至個體中。本文描述的抗CD161抗體之不同投與途徑包括靜脈內、肌肉內、皮下、腹膜內、膀胱內、經皮、脊髓或其他非經腸投與途徑，例如藉由注射或輸注。在一些態樣中，該投與包含非經腸式投與。如本文所用，「非經腸投與」意指除腸內及局部投與之外的投與模式，通常藉由注射，且包括(但不限於)靜脈內、眼內、腹膜內、肌肉內、動脈內、鞘內、淋巴內、病灶內、囊內、眶內、心臟內、皮內、經氣管、皮下、表皮下、關節內、囊下、蛛網膜下、脊柱內、硬膜外及胸骨內注射及輸注以及活體內電穿孔。或者，本文描述之抗體可經由非注射途徑(諸如局部、表皮或黏膜投與途徑，例如經鼻內、經口、經陰道、經直腸、經舌下或局部)投與。投與亦可進行例如一次、複數次及/或歷時一或多個延長之時段。As used herein, "administration" refers to the physical introduction of a composition comprising a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Different routes of administration of the anti-CD161 antibodies described herein include intravenous, intramuscular, subcutaneous, intraperitoneal, intravesical, transdermal, spinal or other non-parenteral routes of administration, such as by injection or infusion. In some aspects, the administration comprises non-parenteral administration. As used herein, "non-parenteral administration" means modes of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intraocular, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. Alternatively, the antibodies described herein may be administered via a non-injection route, such as a topical, epidermal, or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually, or topically. Administration may also be performed, for example, once, multiple times, and/or over one or more extended periods of time.

術語「治療(treat)」、「治療(treating)」及「治療(treatment)」如本文所用係指對該個體進行或投與活性劑至該個體之任何類型之干預或過程，其目標係逆轉、緩解、改善、抑制、或減慢或防止與疾病相關聯之症狀、併發症、病狀或生化指標之進展、發展、嚴重度或復發或增強總存活期。治療可係患有疾病的個體或不患有疾病的個體(例如用於預防)。The terms "treat", "treating" and "treatment" as used herein refer to any type of intervention or process performed on or administered to an individual with the goal of reversing, alleviating, ameliorating, inhibiting, or slowing or preventing the progression, development, severity or recurrence of symptoms, complications, conditions or biochemical markers associated with a disease or enhancing overall survival. Treatment may be of an individual with the disease or an individual without the disease (e.g., for prevention).

術語「有效劑量(effective dose/effective dosage)」係定義為足以達成或至少部分達成所需效應之量。藥物或該治療劑之「治療有效量(therapeutically effective amount)」或「治療有效劑量(therapeutically effective dosage)」係該藥物當單獨使用或與另一治療劑組合使用時促進疾病消退之任何量，該疾病消退藉由以下證明：疾病症狀之嚴重度降低、疾病無症狀期之頻率及持續時間增加、總存活期(自疾病(諸如癌症)之診斷日期或治療開始之時間長度，診斷為患有該疾病的患者仍存活)增加、或由於罹患該疾病所致之受損或失能之預防。藥物之治療有效量或劑量包括「預防有效量」或「預防有效劑量」，其係藥物當單獨或與另一種治療劑組合投與給處於發展出疾病風險或罹患疾病之復發風險中的個體時抑制疾病之發展或復發之任何量。治療劑促進疾病消退或抑制疾病之發展或復發之能力可使用熟練從業人員已知的多種方法來評估，諸如在臨床試驗期間在人類個體中、在預測人類中之功效之動物模型系統中、或藉由分析體外分析中該試劑之活性。The term "effective dose" or "effective dosage" is defined as an amount sufficient to achieve or at least partially achieve a desired effect. A "therapeutically effective amount" or "therapeutically effective dosage" of a drug or therapeutic agent is any amount of the drug that, when used alone or in combination with another therapeutic agent, promotes regression of a disease as evidenced by a decrease in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, an increase in overall survival (the length of time from the date of diagnosis of a disease (such as cancer) or the start of treatment that a patient diagnosed with the disease is still alive), or the prevention of impairment or disability due to having the disease. A therapeutically effective amount or dose of a drug includes a "prophylactically effective amount" or "prophylactically effective dose," which is any amount of a drug that inhibits the development or recurrence of a disease when administered alone or in combination with another therapeutic agent to a subject at risk of developing the disease or at risk of suffering a recurrence of the disease. The ability of a therapeutic agent to promote regression of a disease or inhibit the development or recurrence of a disease can be assessed using a variety of methods known to skilled practitioners, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by analyzing the activity of the agent in in vitro assays.

術語「患者」包括接受預防性或治療性治療的人類及其他哺乳動物個體。The term "patient" includes humans and other mammalian individuals receiving preventive or therapeutic treatment.

如本文所用，術語「個體」包括任何人類動物或非人類的動物。例如，本文描述之方法及組合物可用於治療患有癌症的個體。術語「非人類的動物」包括所有脊椎動物，例如哺乳動物及非哺乳動物，諸如非人類的靈長類動物、綿羊、狗、牛、雞、兩棲動物、爬行動物等。As used herein, the term "subject" includes any human or non-human animal. For example, the methods and compositions described herein can be used to treat a subject suffering from cancer. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

如本文所用，術語「ug」及「uM」可分別與「μg」及「μΜ」互換使用。As used herein, the terms "ug" and "uM" may be used interchangeably with "μg" and "μΜ", respectively.

在以下子章節中更詳細地描述本文揭示之各種態樣。 II.    本發明組合物Various aspects disclosed herein are described in more detail in the following subsections.II. Compositions of the Invention

本發明係關於特異性結合CD161之抗體及其抗原結合部分。在一些態樣中，本文揭示之抗CD161抗體及其抗原結合部分阻斷或減少CD161與CLEC2D之間的相互作用。CD161至CLEC2D之結合抑制T細胞活化。因此，在一些態樣中，本文描述之抗CD161抗體及其抗原結合部分能夠增強人類個體之免疫反應但阻斷或減少CD161-CLEC2D誘導之T細胞活化之抑制。The present invention relates to antibodies and antigen binding portions thereof that specifically bind to CD161. In some aspects, the anti-CD161 antibodies and antigen binding portions thereof disclosed herein block or reduce the interaction between CD161 and CLEC2D. The binding of CD161 to CLEC2D inhibits T cell activation. Therefore, in some aspects, the anti-CD161 antibodies and antigen binding portions thereof described herein can enhance the immune response of a human individual but block or reduce the inhibition of CD161-CLEC2D-induced T cell activation.

任一方法可用於確定抗CD161克服藉由CD161-CLEC2D相互作用介導之T細胞活化抑制之能力。本文描述之抗CD161抗體誘導或增強免疫細胞之細胞技術產生，例如，藉由細胞激素分析確定。在一些態樣中，該細胞激素分析確定自與該抗CD161抗體接觸的免疫細胞分泌的至少一種細胞激素之量，其中該至少一種細胞激素之量之增加指示該抗CD161抗體誘導或增強細胞激素產生。Any method can be used to determine the ability of anti-CD161 to overcome the inhibition of T cell activation mediated by CD161-CLEC2D interaction. The anti-CD161 antibodies described herein induce or enhance the cytokine production of immune cells, for example, as determined by cytokine analysis. In some aspects, the cytokine analysis determines the amount of at least one cytokine secreted from immune cells contacted with the anti-CD161 antibody, wherein an increase in the amount of the at least one cytokine indicates that the anti-CD161 antibody induces or enhances cytokine production.

在一些實施例中，細胞激素產生之增加係至少1.5倍、2倍、3倍、4倍、5倍、6倍、7倍、8倍、9倍或10倍高於對照抗體(例如不結合至CD161之等效抗體同型物，例如不誘導細胞激素產生之抗體)。將該免疫細胞產生之至少一種細胞激素之量與自參考免疫細胞分泌之量進行比較，其中使該參考免疫細胞與對照抗體接觸，且其中自該免疫細胞產生之該至少一種細胞激素之量相對於該參考免疫細胞之增加指示，由於阻斷CD161-CLEC2D相互作用所致之誘導或增強之細胞激素產生。在一些實施例中，該免疫細胞係T細胞，即CD8+或CD4+ T細胞。In some embodiments, the increase in cytokine production is at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold greater than a control antibody (e.g., an equivalent antibody isotype that does not bind to CD161, e.g., an antibody that does not induce cytokine production). The amount of at least one cytokine produced by the immune cell is compared to the amount secreted from a reference immune cell, wherein the reference immune cell is contacted with a control antibody, and wherein an increase in the amount of the at least one cytokine produced by the immune cell relative to the reference immune cell indicates induced or enhanced cytokine production due to blocking CD161-CLEC2D interaction. In some embodiments, the immune cell is a T cell, i.e., a CD8+ or CD4+ T cell.

在一些態樣中，抗CD161抗體及其抗原結合部分誘導或增強免疫細胞之細胞激素產生。在一些態樣中，相較於接觸對照抗體(例如未結合至CD161之等效抗體同型物)或與對照抗體(例如未結合至CD161之等效抗體同型物)接觸之前的免疫細胞，與抗CD161抗體及其抗原結合部分接觸的免疫細胞之細胞激素產生增加至少約1.5倍、至少約2倍、至少約3倍、至少約4倍、至少約5倍、至少約6倍、至少約7倍、至少約8倍、至少約9倍或至少約10倍。在一些態樣中，該免疫細胞係T細胞。在一些態樣中，該T細胞係CD8⁺T細胞或CD4⁺T細胞。在一些態樣中，該免疫細胞係NK細胞。In some aspects, anti-CD161 antibodies and antigen-binding portions thereof induce or enhance cytokine production of immune cells. In some aspects, cytokine production of immune cells contacted with anti-CD161 antibodies and antigen-binding portions thereof increases by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times, compared to immune cells contacted with a control antibody (e.g., an equivalent antibody isotype that is not bound to CD161) or before contact with a control antibody (e.g., an equivalent antibody isotype that is not bound to CD161). In some aspects, the immune cell is a T cell. In some embodiments, the T cell is a CD8⁺ T cell or a CD4⁺ T cell. In some embodiments, the immune cell is a NK cell.

在一些態樣中，使免疫細胞與本文描述之抗CD161抗體及其抗原結合部分接觸導致該免疫細胞之介白素-2 (IL-2)表現增加。在一些態樣中，相較於藉由接觸對照抗體或與對照抗體接觸之前的免疫細胞之IL-2之表現，IL-2表現增加至少約1.5倍、至少約2倍、至少約3倍、至少約4倍、至少約5倍、至少約6倍、至少約7倍、至少約8倍、至少約9倍或至少約10倍。In some aspects, contacting an immune cell with an anti-CD161 antibody and its antigen-binding portion described herein results in an increase in interleukin-2 (IL-2) expression of the immune cell. In some aspects, IL-2 expression is increased by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times compared to the expression of IL-2 of the immune cell before or after contact with a control antibody.

在一些態樣中，使免疫細胞與本文描述之抗CD161抗體及其抗原結合部分接觸導致該免疫細胞之干擾素-γ (IFNg)表現增加。在一些態樣中，相較於藉由接觸對照抗體或與對照抗體接觸之前的免疫細胞之IFNg表現，IFNg表現增加至少約1.5倍、至少約2倍、至少約3倍、至少約4倍、至少約5倍、至少約6倍、至少約7倍、至少約8倍、至少約9倍或至少約10倍。In some aspects, contacting an immune cell with an anti-CD161 antibody and its antigen-binding portion described herein results in an increase in interferon-γ (IFNg) expression by the immune cell. In some aspects, IFNg expression is increased by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times compared to IFNg expression of the immune cell before or after contact with a control antibody.

在一些態樣中，使免疫細胞與本文描述之抗CD161抗體及其抗原結合部分接觸導致該免疫細胞之腫瘤壞死因子-α (TNF-a)表現增加。在一些態樣中，相較於藉由接觸對照抗體或與對照抗體接觸之前的免疫細胞之TNF-a表現，TNF-a表現增加至少約1.5倍、至少約2倍、至少約3倍、至少約4倍、至少約5倍、至少約6倍、至少約7倍、至少約8倍、至少約9倍或至少約10倍。In some aspects, contacting an immune cell with an anti-CD161 antibody and its antigen-binding portion described herein results in an increase in the expression of tumor necrosis factor-alpha (TNF-a) by the immune cell. In some aspects, TNF-a expression is increased by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, or at least about 10 times compared to the expression of TNF-a by the immune cell before or after contact with a control antibody.

在一些態樣中，本文描述之抗CD161抗體及其抗原結合部分以高親和力，例如以10^-6M或更小、10^-7M或更小、10^-8M或更小、10^-9M或更小、10^-10M或更小、10^-11M或更小、10^-12M或更小、10^-12M至10^-7M、10^-11M至10^-7M、10^-10M至10^-7M、或10^-9M至10^-7M之K_D結合至人類CD161。在一些態樣中，本文描述之抗CD161抗體及其抗原結合部分以10^-6M或更小、10^-7M或更小、10^-8M或更小、10^-9M (1 nM)或更小、10^-10M或更小、10^-12M至10^-7M、10^-11M至10^-7M、10^-10M 至10^-7M、10^-9M至10^-7M、或10^-8M至10^-7M之K_D結合至人類CD161，例如，藉由表面電漿子共振，例如，使用Biacore™測定。 II.A.  抗CD161抗體In some aspects, the anti-CD161 antibodies and antigen-binding portions thereof described herein bind to human CD161 with high affinity, for example, with a KD of^10-6 M or less,^10-7 M or less,^10-8 M or less,^10-9 M or less,^10-10 M or less,^10-11 M or less,^10-12 M or less,^10-12 M to^10-7 M,^10-11 M to^10-7 M,^10-10 M to^10-7 M, or^10-9_M to^10-7 M. In some aspects, the anti-CD161 antibodies and antigen-binding portions thereof described herein bind to human CD161 with a KD of^10-6 M or less,^10-7 M or less,^10-8 M or less,^10-9 M (1 nM) or less,^10-10 M or less,^10-12 M to^10-7 M,^10-11 M to^10-7 M,^10-10 M to^10-7 M,^10-9 M to^10-7 M, or^10-8 M to^10-7 M, e.g., as determined by surface_plasmon resonance, e.g., using Biacore™. II.A. Anti-CD161 Antibodies

在一些態樣中，該抗體或其抗原結合部分包含重鏈及輕鏈，其中該重鏈包含重鏈可變區(VH)，且其中該輕鏈包含輕鏈可變區(VL)；其中該VH包含VH互補決定區1 (VH-CDR1)、VH-CDR2及VH-CDR3；其中該VL包含VL-CDR1、VL-CDR2及VL-CDR3；且其中該VH-CDR3包含選自SEQ ID NO: 3、13、23、33、43、53、63、73、83、93、103、113、123、133、143、153、163、173、183、193、203、213、223、233、243、253、263、273、283、293、303、313、323及333所示序列之胺基酸序列。在一些態樣中，該VH-CDR2包含選自SEQ ID NO: 2、12、22、32、42、52、62、72、82、92、102、112、122、132、142、152、162、172、182、192、202、212、222、232、242、252、262、272、282、292、302、312、322及332所示序列之胺基酸序列。在一些態樣中，該VH-CDR1包含選自SEQ ID NO: 1、11、21、31、41、51、61、71、81、91、101、111、121、131、141、151、161、171、181、191、201、211、221、231、241、251、261、271、281、291、301、311、321及331所示序列之胺基酸序列。在一些態樣中，該VL-CDR3包含選自SEQ ID NO: 6、16、26、36、46、56、66、76、86、96、106、116、126、136、146、156、166、176、186、196、206、216、226、236、246、256、266、276、286、296、306、316、326及336所示序列之胺基酸序列。在一些態樣中，該VL-CDR2包含選自SEQ ID NO: 5、15、25、35、45、55、65、75、85、95、105、115、125、135、145、155、165、175、185、195、205、215、225、235、245、255、265、275、285、295、305、315、325及335所示序列之胺基酸序列。在一些態樣中，該VL-CDR1包含選自SEQ ID NO: 4、14、24、34、44、54、64、74、84、94、104、114、124、134、144、154、164、174、184、194、204、214、224、234、244、254、264、274、284、294、304、314、324及334所示序列之胺基酸序列。表2：抗CD161抗體序列In some aspects, the antibody or antigen-binding portion thereof comprises a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region (VH), and wherein the light chain comprises a light chain variable region (VL); wherein the VH comprises a VH complement determining region 1 (VH-CDR1), a VH-CDR2, and a VH-CDR3; wherein the VL comprises a VL-CDR1, a VL-CDR2, and a VL-CDR3; and wherein the VH-CDR3 comprises a sequence selected from SEQ ID NO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, 133, 143, 153, 163, 173, 183, 193, 203, 213, 223, 233, 243, 253, 263, 273, 283, 293, 303, 313, 323 and the amino acid sequence of the sequence shown in 333. In some aspects, the VH-CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, and 332. In some aspects, the VH-CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, and 331. In some aspects, the VL-CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, and 336. In some aspects, the VL-CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325 and 335. In some aspects, the VL-CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324 and 334.Table2: Anti-CD161 Antibody Sequences

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 1所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 2所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 3所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 4所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 5所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 6所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 1所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 2所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 3所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 4所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 5所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 6所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 1所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 2所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 3所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 4所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 5所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 6所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 91所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 92所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 93所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 94所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 95所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 96所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 91, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 92, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 93, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 94, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 95, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 96.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 91所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 92所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 93所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 94所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 95所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 96所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 91, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 92, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 93, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 94, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 95, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 96.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 91所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 92所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 93所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 94所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 95所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 96所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 91, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 92, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 93, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 94, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 95, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 96.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 111所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 112所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 113所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 114所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 115所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 116所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 111, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 112, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 113, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 114, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 116.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 111所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 112所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 113所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 114所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 115所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 116所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 111, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 112, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 113, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 114, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 116.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 111所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 112所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 113所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 114所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 115所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 116所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 111, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 112, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 113, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 114, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 116.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 141所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 142所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 143所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 144所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 145所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 146所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 144, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 145, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 146.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 141所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 142所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 143所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 144所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 145所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 146所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 144, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 145, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 146.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 141所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 142所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 143所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 144所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 145所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 146所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 144, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 145, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 146.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 211所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 212所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 213所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 214所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 215所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 216所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 213, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 216.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 211所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 212所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 213所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 214所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 215所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 216所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 213, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 216.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 211所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 212所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 213所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 214所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 215所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 216所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 213, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 216.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 221所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 222所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 223所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 224所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 225所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 226所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 221, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 222, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 223, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 224, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 225, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 226.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 221所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 222所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 223所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 224所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 225所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 226所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 221, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 222, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 223, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 224, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 225, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 226.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 221所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 222所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 223所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 224所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 225所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 226所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 221, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 222, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 223, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 224, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 225, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 226.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 271所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 272所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 273所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 274所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 275所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 276所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 271, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 272, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 273, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 274, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 275, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 276.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 271所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 272所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 273所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 274所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 275所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 276所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 271, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 272, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 273, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 274, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 275, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 276.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 271所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 272所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 273所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 274所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 275所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 276所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 271, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 272, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 273, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 274, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 275, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 276.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 281所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 282所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 283所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 284所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 285所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 286所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 281, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 282, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 283, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 284, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 285, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 286.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 281所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 282所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 283所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 284所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 285所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 286所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 281, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 282, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 283, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 284, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 285, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 286.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 281所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 282所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 283所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 284所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 285所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 286所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 281, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 282, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 283, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 284, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 285, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 286.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 291所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 292所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 293所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 294所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 295所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 296所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 291, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 292, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 293, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 294, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 295, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 296.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 291所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 292所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 293所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 294所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 295所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 296所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 291, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 292, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 293, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 294, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 295, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 296.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 291所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 292所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 293所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 294所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 295所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 296所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 291, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 292, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 293, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 294, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 295, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 296.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 301所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 302所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 303所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 304所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 305所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 306所示胺基酸序列之VL-CDR3。In some aspects, the antibody or its antigen-binding portion contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 302, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 303, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 306.

在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 301所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 302所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 303所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 304所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 305所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 306所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 302, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 303, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 306.

在一些態樣中，該抗體或其抗原結合部分結合人類CD161上與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 301所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 302所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 303所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 304所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 305所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 306所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof binds to the same antigenic determinant on human CD161 as a reference antibody, wherein the reference antibody contains a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 302, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 303, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 306.

在一些態樣中，該抗體或其抗原結合部分含有包含與選自SEQ ID NO: 7、17、27、37、47、57、67、77、87、97、107、117、127、137、147、157、167、177、187、197、207、217、227、237、247、257、267、277、287、297、307、317、327及337之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 7所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 97所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 117所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 147所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 217所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 227所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有VH包含與SEQ ID NO: 277所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 287所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有與SEQ ID NO: 297所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 307所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH。In some aspects, the antibody, or antigen binding portion thereof, contains a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, and 337. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 7. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 97. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 117. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 147. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 217. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 227. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 277. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 287. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 297. In some aspects, the antibody, or antigen-binding portion thereof, contains a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 307.

在一些態樣中，該抗體或其抗原結合部分含有包含與選自SEQ ID NO: 8、18、28、38、48、58、68、78、88、98、108、118、128、138、148、158、168、178、188、198、208、218、228、238、248、258、268、278、288、298、308、318、328及338之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 8所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 98所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 118所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 148所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 218所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 228所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 278所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 288所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 298所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含與SEQ ID NO: 308所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody, or antigen binding portion thereof, contains a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 8. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 98. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 118. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 148. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 218. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 228. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 278. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 288. In some aspects, the antibody, or antigen-binding portion thereof, comprises a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 298. In some aspects, the antibody, or antigen-binding portion thereof, contains a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 308.

在一些態樣中，該抗體或其抗原結合部分含有包含選自SEQ ID NO: 7、17、27、37、47、57、67、77、87、97、107、117、127、137、147、157、167、177、187、197、207、217、227、237、247、257、267、277、287、297、307、317、327及337之胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 7所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 97所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 117所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 147所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 217所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 227所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 277所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 287所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 297所示胺基酸序列之VH。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 307所示胺基酸序列之VH。In some aspects, the antibody, or an antigen-binding portion thereof, comprises a VH comprising an amino acid sequence selected from SEQ ID NO: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, and 337. In some aspects, the antibody, or an antigen-binding portion thereof, comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 7. In some aspects, the antibody, or an antigen-binding portion thereof, comprises a VH comprising an amino acid sequence set forth in SEQ ID NO: 97. In some aspects, the antibody or its antigen-binding portion thereof contains a VH comprising the amino acid sequence of SEQ ID NO: 117. In some aspects, the antibody or its antigen-binding portion thereof contains a VH comprising the amino acid sequence of SEQ ID NO: 147. In some aspects, the antibody or its antigen-binding portion thereof contains a VH comprising the amino acid sequence of SEQ ID NO: 217. In some aspects, the antibody or its antigen-binding portion thereof contains a VH comprising the amino acid sequence of SEQ ID NO: 227. In some aspects, the antibody or its antigen-binding portion thereof contains a VH comprising the amino acid sequence of SEQ ID NO: 277. In some aspects, the antibody or its antigen-binding portion thereof contains a VH comprising the amino acid sequence of SEQ ID NO: 287. In some aspects, the antibody or its antigen-binding portion thereof contains a VH comprising the amino acid sequence of SEQ ID NO: 297. In some aspects, the antibody or antigen-binding portion thereof comprises a VH comprising the amino acid sequence shown in SEQ ID NO: 307.

在一些態樣中，該抗體或其抗原結合部分含有包含選自SEQ ID NO: 8、18、28、38、48、58、68、78、88、98、108、118、128、138、148、158、168、178、188、198、208、218、228、238、248、258、268、278、288、298、308、318、328及338之胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 8所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 98所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 118所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 148所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 218所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 228所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 278所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 288所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 298所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 308所示胺基酸序列之VL。In some aspects, the antibody, or an antigen binding portion thereof, comprises a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338. In some aspects, the antibody, or an antigen binding portion thereof, comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 8. In some aspects, the antibody, or an antigen binding portion thereof, comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 98. In some aspects, the antibody or its antigen-binding portion thereof contains a VL comprising the amino acid sequence set forth in SEQ ID NO: 118. In some aspects, the antibody or its antigen-binding portion thereof contains a VL comprising the amino acid sequence set forth in SEQ ID NO: 148. In some aspects, the antibody or its antigen-binding portion thereof contains a VL comprising the amino acid sequence set forth in SEQ ID NO: 218. In some aspects, the antibody or its antigen-binding portion thereof contains a VL comprising the amino acid sequence set forth in SEQ ID NO: 228. In some aspects, the antibody or its antigen-binding portion thereof contains a VL comprising the amino acid sequence set forth in SEQ ID NO: 278. In some aspects, the antibody or its antigen-binding portion thereof contains a VL comprising the amino acid sequence set forth in SEQ ID NO: 288. In some aspects, the antibody or its antigen-binding portion thereof contains a VL comprising the amino acid sequence set forth in SEQ ID NO: 298. In some aspects, the antibody or antigen-binding portion thereof comprises a VL comprising the amino acid sequence shown in SEQ ID NO: 308.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 7所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 8所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 7; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 8.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 7所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 8所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 1所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 2所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 3所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 4所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 5所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 6所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 7; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 8; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL comprising the amino acid sequence of SEQ ID NO: 4 NO: 4, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 6.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 7所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 8所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 1所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 2所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 3所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 4所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 5所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 6所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 7; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 8; wherein the antibody or antigen-binding portion thereof cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VL comprising the amino acid sequence of SEQ ID NO: 3, a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 3, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 6.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 7所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 8所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 1所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 2所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 3所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 4所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 5所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 6所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or an antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 7; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 8; wherein the antibody, or an antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 6 VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, and VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 6.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 97所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 98所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 97; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 98.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 97所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 98所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 91所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 92所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 93所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 94所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 95所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 96所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 97; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 98; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 91, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 92, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 93, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 94, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 95, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 96.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 97所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 98所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 91所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 92所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 93所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 94所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 95所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 96所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 97; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 98; wherein the antibody, or antigen-binding portion thereof, cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 91, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 92, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 93, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 94, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 95, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 96.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 97所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 98所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 91所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 92所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 93所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 94所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 95所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 96所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 97; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 98; wherein the antibody, or antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 91, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 92, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 93, a VL comprising the amino acid sequence of SEQ ID NO: The present invention further comprises a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 94, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 95, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 96.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 117所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 118所示胺基酸序列之具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 117; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 118.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 117所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 118所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 111所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 112所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 113所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 114所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 115所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 116所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 117; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 118; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 111, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 112, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 113, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 114, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 115, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 116.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 117所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 118所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 111所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 112所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 113所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 114所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 115所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 116所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 117; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 118; wherein the antibody, or antigen-binding portion thereof, cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 111, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 112, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 113, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 114, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 115, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 116.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 117所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 118所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 111所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 112所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 113所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 114所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 115所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 116所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 117; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 118; wherein the antibody, or antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 111, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 112, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 113, a VL comprising the amino acid sequence of SEQ ID NO: The present invention further comprises a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 114, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 115, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 116.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 147所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 148所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 147; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 148.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 147所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 148所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 141所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 142所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 143所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 144所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 145所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 146所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 147; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 148; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142, a VL comprising the amino acid sequence of SEQ ID NO: The invention also comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 143, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 144, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 145, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 146.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 147所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 148所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 141所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 142所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 143所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 144所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 145所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 146所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 147; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 148; wherein the antibody, or antigen-binding portion thereof, cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142, a VL comprising the amino acid sequence of SEQ ID NO: The invention also comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 143, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 144, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 145, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 146.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 147所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 148所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 141所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 142所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 143所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 144所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 145所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 146所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 147; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 148; wherein the antibody, or antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143, a VL comprising the amino acid sequence of SEQ ID NO: The present invention relates to a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 144, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 145, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 146.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 217所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 218所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 217; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 218.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 217所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 218所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 211所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 212所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 213所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 214所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 215所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 216所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 217; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 218; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 213, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 216.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 217所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 218所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 211所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 212所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 213所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 214所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 215所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 216所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 217; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 218; wherein the antibody, or antigen-binding portion thereof, cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 213, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 216.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 217所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 218所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 211所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 212所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 213所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 214所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 215所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 216所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 217; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 218; wherein the antibody, or antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 213, a VL comprising the amino acid sequence of SEQ ID NO: The present invention further comprises a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 216.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 227所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 228所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 227; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 228.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 227所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 228所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 221所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 222所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 223所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 224所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 225所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 226所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 227; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 228; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 221, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 222, a VL comprising the amino acid sequence of SEQ ID NO: The present invention relates to a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 223, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 224, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 225, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 226.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 227所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 228所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 221所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 222所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 223所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 224所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 225所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 226所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 227; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 228; wherein the antibody, or antigen-binding portion thereof, cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 221, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 222, a VL comprising the amino acid sequence of SEQ ID NO: The present invention relates to a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 223, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 224, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 225, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 226.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 227所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 228所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 221所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 222所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 223所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 224所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 225所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 226所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 227; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 228; wherein the antibody, or antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 221, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 222, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 223, a VL comprising the amino acid sequence of SEQ ID NO: The present invention further comprises a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 224, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 225, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 226.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 277所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 278所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 277; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 278.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 277所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 278所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 271所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 272所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 273所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 274所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 275所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 276所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 277; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 278; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 271, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 272, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 273, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 274, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 275, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 276.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 277所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 278所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 271所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 272所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 273所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 274所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 275所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 276所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 277; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 278; wherein the antibody, or antigen-binding portion thereof, cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 271, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 272, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 273, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 274, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 275, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 276.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 277所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 278所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 271所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 272所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 273所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 274所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 275所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 276所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 277; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 278; wherein the antibody, or antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 271, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 272, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 273, a VL comprising the amino acid sequence of SEQ ID NO: The present invention relates to a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 274, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 275, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 276.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 287所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 288所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 287; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 288.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 287所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 288所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 281所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 282所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 283所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 284所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 285所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 286所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 287; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 288; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 281, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 282, a VL comprising the amino acid sequence of SEQ ID NO: The invention also comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 283, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 284, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 285, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 286.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 287所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 288所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 281所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 282所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 283所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 284所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 285所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 286所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 287; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 288; wherein the antibody, or antigen-binding portion thereof, cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 281, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 282, a VL comprising the amino acid sequence of SEQ ID NO: The invention also comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 283, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 284, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 285, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 286.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 287所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 288所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 281所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 282所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 283所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 284所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 285所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 286所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 287; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 288; wherein the antibody, or antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 281, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 282, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 283, a VL comprising the amino acid sequence of SEQ ID NO: The present invention further comprises a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 284, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 285, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 286.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 297所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 298所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 297; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 298.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 297所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 298所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 291所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 292所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 293所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 294所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 295所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 296所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 297; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 298; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 291, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 292, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 293, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 294, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 295, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 296.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 297所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 298所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 291所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 292所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 293所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 294所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 295所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 296所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 297; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 298; wherein the antibody, or antigen-binding portion thereof, cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 291, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 292, a VL comprising the amino acid sequence of SEQ ID NO: The invention further comprises a VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 293, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 294, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 295, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 296.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 297所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 298所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 291所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 292所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 293所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 294所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 295所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 296所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 297; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 298; wherein the antibody, or antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 291, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 292, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 293, a VL comprising the amino acid sequence of SEQ ID NO: The present invention further comprises a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 294, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 295, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 296.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 307所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 308所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL。In some aspects, the antibody or antigen-binding portion thereof contains (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 307; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 308.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 307所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 308所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 301所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 302所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 303所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 304所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 305所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 306所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 307; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 308; wherein the antibody, or antigen-binding portion thereof, binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 302, a VL comprising the amino acid sequence of SEQ ID NO: A VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 303, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 304, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 305, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 306.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 307所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 308所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 301所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 302所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 303所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 304所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 305所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 306所示胺基酸序列之VL-CDR3。In some aspects, the antibody or antigen-binding portion thereof comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 307; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 308; wherein the antibody or antigen-binding portion thereof cross-competitively binds to human CD161 with a reference antibody, wherein the reference antibody comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 302, a VL comprising the amino acid sequence of SEQ ID NO: A VH-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 303, a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 304, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 305, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 306.

在一些態樣中，該抗體或其抗原結合部分含有(i)包含與SEQ ID NO: 307所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VH；及(ii)包含與SEQ ID NO: 308所示胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列之VL；其中該抗體或其抗原結合部分含有包含SEQ ID NO: 301所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 302所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 303所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 304所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 305所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 306所示胺基酸序列之VL-CDR3。In some aspects, the antibody, or an antigen-binding portion thereof, comprises (i) a VH comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 307; and (ii) a VL comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 308; wherein the antibody, or an antigen-binding portion thereof, comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 302, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 303, a VL comprising the amino acid sequence of SEQ ID NO: The present invention further comprises a VL-CDR1 comprising the amino acid sequence shown in SEQ ID NO: 304, a VL-CDR2 comprising the amino acid sequence shown in SEQ ID NO: 305, and a VL-CDR3 comprising the amino acid sequence shown in SEQ ID NO: 306.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 7所示胺基酸序列之VH、及包含SEQ ID NO: 8所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 7所示胺基酸序列之VH、及包含SEQ ID NO: 8所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 7所示胺基酸序列之VH、及包含SEQ ID NO: 8所示胺基酸序列之VL。In some aspects, the antibody or its antigen-binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 7, and a VL comprising the amino acid sequence of SEQ ID NO: 8. In some aspects, the antibody or its antigen-binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 7, and a VL comprising the amino acid sequence of SEQ ID NO: 8. In some aspects, the antibody or its antigen-binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 7, and a VL comprising the amino acid sequence of SEQ ID NO: 8.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 97所示胺基酸序列之VH、及包含SEQ ID NO: 98所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 97所示胺基酸序列之VH、及包含SEQ ID NO: 98所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 97所示胺基酸序列之VH、及包含SEQ ID NO: 98所示胺基酸序列之VL。In some aspects, the antibody or its antigen binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 97, and a VL comprising the amino acid sequence of SEQ ID NO: 98. In some aspects, the antibody or its antigen binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 97, and a VL comprising the amino acid sequence of SEQ ID NO: 98. In some aspects, the antibody or its antigen binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 97, and a VL comprising the amino acid sequence of SEQ ID NO: 98.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 117所示胺基酸序列之VH、及包含SEQ ID NO: 118所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 117所示胺基酸序列之VH、及包含SEQ ID NO: 118所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 117所示胺基酸序列之VH、及包含SEQ ID NO: 118所示胺基酸序列之VL。In some aspects, the antibody or its antigen-binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 117, and a VL comprising the amino acid sequence of SEQ ID NO: 118. In some aspects, the antibody or its antigen-binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 117, and a VL comprising the amino acid sequence of SEQ ID NO: 118. In some aspects, the antibody or its antigen-binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 117, and a VL comprising the amino acid sequence of SEQ ID NO: 118.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 147所示胺基酸序列之VH、及包含SEQ ID NO: 148所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 147所示胺基酸序列之VH、及包含SEQ ID NO: 148所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 147所示胺基酸序列之VH、及包含SEQ ID NO: 148所示胺基酸序列之VL。In some aspects, the antibody or its antigen-binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 147, and a VL comprising the amino acid sequence of SEQ ID NO: 148. In some aspects, the antibody or its antigen-binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 147, and a VL comprising the amino acid sequence of SEQ ID NO: 148. In some aspects, the antibody or its antigen-binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 147, and a VL comprising the amino acid sequence of SEQ ID NO: 148.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 217所示胺基酸序列之VH、及包含SEQ ID NO: 218所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 217所示胺基酸序列之VH、及包含SEQ ID NO: 218所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 217所示胺基酸序列之VH、及包含SEQ ID NO: 218所示胺基酸序列之VL。In some aspects, the antibody or its antigen binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 217, and a VL comprising the amino acid sequence of SEQ ID NO: 218. In some aspects, the antibody or its antigen binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 217, and a VL comprising the amino acid sequence of SEQ ID NO: 218. In some aspects, the antibody or its antigen binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 217, and a VL comprising the amino acid sequence of SEQ ID NO: 218.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 227所示胺基酸序列之VH、及包含SEQ ID NO: 228所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 227所示胺基酸序列之VH、及包含SEQ ID NO: 228所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 227所示胺基酸序列之VH、及包含SEQ ID NO: 228所示胺基酸序列之VL。In some aspects, the antibody or its antigen-binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 227, and a VL comprising the amino acid sequence of SEQ ID NO: 228. In some aspects, the antibody or its antigen-binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 227, and a VL comprising the amino acid sequence of SEQ ID NO: 228. In some aspects, the antibody or its antigen-binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 227, and a VL comprising the amino acid sequence of SEQ ID NO: 228.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 277所示胺基酸序列之VH、及包含SEQ ID NO: 278所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 277所示胺基酸序列之VH、及包含SEQ ID NO: 278所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 277所示胺基酸序列之VH、及包含SEQ ID NO: 278所示胺基酸序列之VL。In some aspects, the antibody or its antigen binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 277, and a VL comprising the amino acid sequence of SEQ ID NO: 278. In some aspects, the antibody or its antigen binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 277, and a VL comprising the amino acid sequence of SEQ ID NO: 278. In some aspects, the antibody or its antigen binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 277, and a VL comprising the amino acid sequence of SEQ ID NO: 278.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 287所示胺基酸序列之VH、及包含SEQ ID NO: 288所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 287所示胺基酸序列之VH、及包含SEQ ID NO: 288所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 287所示胺基酸序列之VH、及包含SEQ ID NO: 288所示胺基酸序列之VL。In some aspects, the antibody or its antigen binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 287, and a VL comprising the amino acid sequence of SEQ ID NO: 288. In some aspects, the antibody or its antigen binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 287, and a VL comprising the amino acid sequence of SEQ ID NO: 288. In some aspects, the antibody or its antigen binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 287, and a VL comprising the amino acid sequence of SEQ ID NO: 288.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 297所示胺基酸序列之VH、及包含SEQ ID NO: 298所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 297所示胺基酸序列之VH、及包含SEQ ID NO: 298所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 297所示胺基酸序列之VH、及包含SEQ ID NO: 298所示胺基酸序列之VL。In some aspects, the antibody or its antigen binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 297, and a VL comprising the amino acid sequence of SEQ ID NO: 298. In some aspects, the antibody or its antigen binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 297, and a VL comprising the amino acid sequence of SEQ ID NO: 298. In some aspects, the antibody or its antigen binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 297, and a VL comprising the amino acid sequence of SEQ ID NO: 298.

在一些態樣中，該抗體或其抗原結合部分含有包含SEQ ID NO: 307所示胺基酸序列之VH、及包含SEQ ID NO: 308所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分結合與參考抗體相同的抗原決定基，其中該參考抗體含有包含SEQ ID NO: 307所示胺基酸序列之VH、及包含SEQ ID NO: 308所示胺基酸序列之VL。在一些態樣中，該抗體或其抗原結合部分與參考抗體交叉競爭結合至人類CD161，其中該參考抗體含有包含SEQ ID NO: 307所示胺基酸序列之VH、及包含SEQ ID NO: 308所示胺基酸序列之VL。In some aspects, the antibody or its antigen-binding portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 307, and a VL comprising the amino acid sequence of SEQ ID NO: 308. In some aspects, the antibody or its antigen-binding portion binds to the same antigenic determinant as a reference antibody, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 307, and a VL comprising the amino acid sequence of SEQ ID NO: 308. In some aspects, the antibody or its antigen-binding portion cross-competes with a reference antibody for binding to human CD161, wherein the reference antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 307, and a VL comprising the amino acid sequence of SEQ ID NO: 308.

在一些態樣中，該抗體或其抗原結合部分包含一或多個轉譯後修飾。在一些態樣中，該抗體或其抗原結合部分包含一或多個轉譯後修飾，其增加該抗體或其抗原結合部分之活體內半衰期。在一些態樣中，該抗體或其抗原結合部分經聚乙二醇化。In some aspects, the antibody or its antigen-binding portion comprises one or more post-translational modifications. In some aspects, the antibody or its antigen-binding portion comprises one or more post-translational modifications that increase the in vivo half-life of the antibody or its antigen-binding portion. In some aspects, the antibody or its antigen-binding portion is pegylated.

在一些態樣中，該抗體之該抗原結合部分包含VHH、vNAR、微體、奈米抗體、scFv或其任何組合。 II.B.   雙特異性及多特異性抗體In some embodiments, the antigen-binding portion of the antibody comprises a VHH, a vNAR, a microbody, a nanobody, a scFv, or any combination thereof.II.B.   Bispecific and multispecific antibodies

本文所述的抗CD161抗體及其抗原結合部分可用於形成雙特異性及多特異性分子。本文所述的抗CD161抗體或其抗原結合部分可經衍生或連接至另一功能分子，例如另一肽或蛋白質(例如受體之另一抗體或配位體)以產生結合至至少兩個不同結合位點或靶分子之雙特異性分子。例如，抗CD161抗體可連接至特異性結合至腫瘤抗原之抗體或scFv。本文所述的抗體可事實上衍生或連接至多於一個其他功能分子以產生結合至多於兩個不同結合位點及/或靶分子之多特異性分子。為了產生本文所述的雙特異性分子，本文所述的抗體可功能上連接(例如，藉由化學偶聯、遺傳融合、非共價締合或其他方式)至一或多個其他結合分子，諸如另一抗體、抗體片段、肽或結合模擬物，使得雙特異性分子產生。The anti-CD161 antibodies and their antigen binding portions described herein can be used to form bispecific and multispecific molecules. The anti-CD161 antibodies or their antigen binding portions described herein can be derivatized or linked to another functional molecule, such as another peptide or protein (e.g., another antibody or ligand of a receptor) to produce bispecific molecules that bind to at least two different binding sites or target molecules. For example, anti-CD161 antibodies can be linked to antibodies or scFvs that specifically bind to tumor antigens. The antibodies described herein can actually be derived or linked to more than one other functional molecule to produce multispecific molecules that bind to more than two different binding sites and/or target molecules. To generate the bispecific molecules described herein, the antibodies described herein can be functionally linked (e.g., by chemical coupling, genetic fusion, non-covalent association or other means) to one or more other binding molecules, such as another antibody, antibody fragment, peptide or binding mimetic, so that a bispecific molecule is generated.

因此，本文提供包含針對CD161之至少一種第一結合特異性及針對第二靶抗原決定基之第二結合特異性之雙特異性分子。本文進一步提供包含針對CD161之至少一種第一結合特異性、針對第二靶抗原決定基之第二結合特異性、及針對第三靶抗原決定基之第三結合特異性之多特異性分子。Thus, provided herein are bispecific molecules comprising at least one first binding specificity for CD161 and a second binding specificity for a second target antigenic determinant. Further provided herein are multispecific molecules comprising at least one first binding specificity for CD161, a second binding specificity for a second target antigenic determinant, and a third binding specificity for a third target antigenic determinant.

在一些態樣中，本文所述之雙特異性及多特異性分子包含作為結合特異性之至少一種抗體、或其抗體片段，包括例如Fab、Fab'、F(ab')2、Fv或單鏈Fv (scFv)。該抗體亦可係輕鏈或重鏈二聚體、或其任何最小片段，諸如Fv或單鏈構築體，如Ladner等人美國專利第4,946,778號中所述。In some embodiments, the bispecific and multispecific molecules described herein include at least one antibody or antibody fragment thereof as a binding specificity, including, for example, Fab, Fab', F(ab')2, Fv or single-chain Fv (scFv). The antibody may also be a light chain or heavy chain dimer, or any minimal fragment thereof, such as an Fv or single-chain construct, as described in Ladner et al., U.S. Patent No. 4,946,778.

雖然較佳為人類單株抗體，但可用於本文所述之雙特異性及多特異性分子中之其他抗體係鼠類、嵌合及人類化單株抗體。Although human monoclonal antibodies are preferred, other antibodies that can be used in the bispecific and multispecific molecules described herein are murine, chimeric, and humanized monoclonal antibodies.

本文所述之雙特異性及多特異性分子可藉由使用此項技術中已知的方法結合成分結合特異性來製備。例如，可分開產生雙特異性或多特異性分子之每種結合特異性且然後彼此結合。當結合特異性為蛋白質或肽時，可使用多種偶聯劑或交聯劑進行共價結合。交聯劑之實例包括蛋白A、碳二亞胺、N-琥珀醯亞胺基-S-乙醯基-硫代乙酸酯(SATA)、5,5'-二硫雙(2-硝基苯甲酸) (DTNB)、鄰-伸苯基二馬來醯亞胺(oPDM)、N-琥珀醯亞胺基-3-(2-吡啶基二硫)丙酸酯(SPDP)、及4-(N-馬來醯亞胺基甲基)環己烷-1-甲酸磺基琥珀醯亞胺酯(磺基-SMCC) (參見，例如，Karpovsky等人(1984)J. Exp. Med. 160: 1686；Liu, MA等人(1985)Proc. Natl. Acad. Sci. USA82:8648)。其他方法包括彼等描述於Paulus (1985)Behring Ins. Mitt. No. 78, 118-132；Brennan等人(1985)Science229:81-83)、及Glennie等人(1987)J. Immunol. 139: 2367-2375)中者。一些結合劑係SATA及磺基-SMCC，二者均可自Pierce Chemical Co. (Rockford, IL)獲得。The bispecific and multispecific molecules described herein can be prepared by combining the component binding specificities using methods known in the art. For example, each binding specificity of the bispecific or multispecific molecule can be generated separately and then combined with each other. When the binding specificity is a protein or peptide, a variety of coupling agents or cross-linking agents can be used for covalent conjugation. Examples of cross-linking agents include protein A, carbodiimide, N-succinimidyl-S-acetyl-thioacetate (SATA), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), and sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC) (see, e.g., Karpovsky et al. (1984)J. Exp. Med . 160: 1686; Liu, MA et al. (1985)Proc. Natl. Acad. Sci. USA 82:8648). Other methods include those described in Paulus (1985)Behring Ins. Mitt . No. 78, 118-132; Brennan et al. (1985)Science 229:81-83), and Glennie et al. (1987)J. Immunol . 139:2367-2375). Some binding agents are SATA and sulfo-SMCC, both available from Pierce Chemical Co. (Rockford, IL).

當結合特異性為抗體時，其可經由兩個重鏈之C端鉸鏈區之硫氫基鍵結而結合。在一些態樣中，鉸鏈區經修飾以在結合前含有奇數個硫氫基殘基(較佳一個)。When the binding specificity is an antibody, it can be bound via sulfhydryl bonding of the C-terminal hinge regions of the two heavy chains. In some embodiments, the hinge region is modified to contain an odd number of sulfhydryl residues (preferably one) before binding.

或者，多種結合特異性可在相同載體中編碼且在相同宿主細胞中表現且組裝。此方法特別有用，其中雙特異性分子係mAb x mAb、mAb x Fab、mAb x (scFv)₂、Fab x F(ab')₂或配位體 x Fab融合蛋白。雙特異性抗體可包含在每個重鏈的C端包含scFv之抗體。本文所述的雙特異性分子可係包含一個單鏈抗體及結合決定子之單鏈分子、或包含兩個結合決定子之單鏈雙特異性分子。雙特異性分子可包含至少兩個單鏈分子。用於製備雙特異性分子之方法描述於例如美國專利第5,260,203號；美國專利第5,455,030號；美國專利第4,881,175號；美國專利第5,132,405號；美國專利第5,091,513號；美國專利第5,476,786號；美國專利第5,013,653號；美國專利第5,258,498號；及美國專利第5,482,858號中。Alternatively, multiple binding specificities may be encoded in the same vector and expressed and assembled in the same host cell. This method is particularly useful where the bispecific molecule is a mAb x mAb, mAb x Fab, mAb x (scFv)₂ , Fab x F(ab')₂ , or ligand x Fab fusion protein. The bispecific antibody may comprise an antibody comprising an scFv at the C-terminus of each heavy chain. The bispecific molecule described herein may be a single chain molecule comprising a single chain antibody and a binding determinant, or a single chain bispecific molecule comprising two binding determinants. The bispecific molecule may comprise at least two single chain molecules. Methods for making bispecific molecules are described, for example, in U.S. Patent No. 5,260,203; U.S. Patent No. 5,455,030; U.S. Patent No. 4,881,175; U.S. Patent No. 5,132,405; U.S. Patent No. 5,091,513; U.S. Patent No. 5,476,786; U.S. Patent No. 5,013,653; U.S. Patent No. 5,258,498; and U.S. Patent No. 5,482,858.

雙特異性及多特異性分子對其特定靶之結合可使用技術公認的方法(諸如酶聯免疫吸附分析(ELISA)、放射免疫分析(RIA)、FACS分析、生物分析(例如生長抑制)或西方墨點分析(Western Blot assay))來確認。此等分析中之各者一般藉由採用對所關注複合物具有特異性之經標記之試劑(例如抗體)偵測特別關注的蛋白質-抗體複合物之存在。Binding of bispecific and multispecific molecules to their specific targets can be confirmed using art-recognized methods such as enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis, bioassays (e.g., growth inhibition), or Western Blot assays. Each of these assays generally detects the presence of a protein-antibody complex of particular interest by employing a labeled reagent (e.g., an antibody) specific for the complex of interest.

在一些態樣中，該雙特異性抗體包含(i)本文揭示之抗CD161抗體或其抗原結合部分及(ii)特異性結合腫瘤抗原之抗體或其抗原結合部分。在一些態樣中，該腫瘤抗原選自CD19、TRAC、TCRβ、BCMA、CLL-1、CS1、CD38、CD19、TSHR、CD123、CD22、CD30、CD70、CD171、CD33、EGFRvIII、GD2、GD3、Tn Ag、PSMA、ROR1、ROR2、GPC1、GPC2、FLT3、FAP、TAG72、CD44v6、CEA、EPCAM、B7H3、KIT、IL-13Ra2、間皮素、IL-l lRa、PSCA、PRSS21、VEGFR2、LewisY、CD24、PDGFR-β、SSEA-4、CD20、葉酸受體α、ERBB2 (Her2/neu)、MUC1、MUC16、EGFR、NCAM、前列腺酶(prostase)、PAP、ELF2M、艾普瑞林(Ephrin) B2、IGF-I受體、CAIX、LMP2、gplOO、bcr-abl、酪胺酸酶、EphA2、岩藻糖基GM1、sLe、GM3、TGS5、HMWMAA、鄰-乙醯基-GD2、葉酸受體β、TEM1/CD248、TEM7R、CLDN6、GPRC5D、CXORF61、CD97、CD179a、ALK、聚唾液酸、PLAC1、GloboH、NY-BR-1、UPK2、HAVCR1、ADRB3、PANX3、GPR20、LY6K、OR51E2、TARP、WTl、NY-ESO-1、LAGE-la、MAGE-Al、豆莢蛋白(legumain)、HPV E6、E7、MAGE Al、ETV6-AML、精子蛋白17、XAGE1、Tie 2、MAD-CT-1、MAD-CT-2、Fos相關抗原1、p53、p53突變體、普洛斯坦、存活素(surviving)、端粒酶、PCTA-1/半乳糖凝集素8、MelanA/MARTl、Ras突變體、hTERT、肉瘤易位斷點、ML-IAP、ERG (TMPRSS2 ETS融合基因)、NA17、PAX3、雄激素受體、周期蛋白Bl、MYCN、RhoC、TRP-2、CYP1B1、BORIS、SART3、PAX5、OY-TES1、LCK、AKAP-4、SSX2、RAGE-1、人類端粒酶逆轉錄酶、RU1、RU2、腸道羧基酯酶、mut hsp70-2、CD79a、CD79b、CD72、LAIR1、FCAR、LILRA2、CD300LF、CLEC12A、BST2、EMR2、LY75、GPC3、FCRL5、IGLL1、CD2、CD3ε、CD4、CD5、CD7、APRIL蛋白之細胞外部分或其任何組合。在一些態樣中，TCR靶向AFP、CD19、TRAC、TCRβ、BCMA、CLL-1、CS1、CD38、CD19、TSHR、CD123、CD22、CD30、CD171、CD33、EGFRvIII、GD2、GD3、Tn Ag、PSMA、ROR1、ROR2、GPC1、GPC2、FLT3、FAP、TAG72、CD44v6、CEA、EPCAM、B7H3、KIT、IL-13Ra2、間皮素、IL-l lRa、PSCA、PRSS21、VEGFR2、LewisY、CD24、PDGFR-β、SSEA-4、CD20、葉酸受體α、ERBB2 (Her2/neu)、MUC1、MUC16、EGFR、NCAM、前列腺酶、PAP、ELF2M、艾普瑞林B2、IGF-I受體、CAIX、LMP2、gplOO、bcr-abl、酪胺酸酶、EphA2、岩藻糖基GM1、sLe、GM3、TGS5、HMWMAA、鄰-乙醯基-GD2、葉酸受體β、TEM1/CD248、TEM7R、CLDN6、GPRC5D、CXORF61、CD97、CD179a、ALK、聚唾液酸、PLAC1、GloboH、NY-BR-1、UPK2、HAVCR1、ADRB3、PANX3、GPR20、LY6K、OR51E2、TARP、WTl、NY-ESO-1、LAGE-la、MAGE-Al、豆莢蛋白、HPV E6、E7、MAGE Al、ETV6-AML、精子蛋白17、XAGE1、Tie 2、MAD-CT-1、MAD-CT-2、Fos相關抗原1、p53、p53突變體、普洛斯坦、存活素、端粒酶、PCTA-1/半乳糖凝集素8、MelanA/MARTl、Ras突變體、hTERT、肉瘤易位斷點、ML-IAP、ERG (TMPRSS2 ETS融合基因)、NA17、PAX3、雄激素受體、周期蛋白Bl、MYCN、RhoC、TRP-2、CYP1B1、BORIS、SART3、PAX5、OY-TES1、LCK、AKAP-4、SSX2、RAGE-1、人類端粒酶逆轉錄酶、RU1、RU2、腸道羧基酯酶、mut hsp70-2、CD79a、CD79b、CD72、LAIR1、FCAR、LILRA2、CD300LF、CLEC12A、BST2、EMR2、LY75、GPC3、FCRL5、IGLL1、CD2、CD3ε、CD4、CD5、CD7、APRIL蛋白之細胞外部分及其任何組合。在一些態樣中，該雙特異性抗體包含(i)本文揭示之抗CD161抗體或其抗原結合部分及(ii)特異性結合選自下列之腫瘤抗原之抗體或其抗原結合部分：CD19、TRAC、TCRβ、BCMA、CLL-1、CS1、CD38、CD19、TSHR、CD123、CD22、CD30、CD70、CD171、CD33、EGFRvIII、GD2、GD3、Tn Ag、PSMA、ROR1、ROR2、GPC1、GPC2、FLT3、FAP、TAG72、CD44v6、CEA、EPCAM、B7H3、KIT、IL-13Ra2、間皮素、IL-l lRa、PSCA、PRSS21、VEGFR2、LewisY、CD24、PDGFR-β、SSEA-4、CD20、葉酸受體α、ERBB2 (Her2/neu)、MUC1、MUC16、EGFR、NCAM、前列腺酶、PAP、ELF2M、艾普瑞林B2、IGF-I受體、CAIX、LMP2、gplOO、bcr-abl、酪胺酸酶、EphA2、岩藻糖基GM1、sLe、GM3、TGS5、HMWMAA、鄰-乙醯基-GD2、葉酸受體β、TEM1/CD248、TEM7R、CLDN6、GPRC5D、CXORF61、CD97、CD179a、ALK、聚唾液酸、PLAC1、GloboH、NY-BR-1、UPK2、HAVCR1、ADRB3、PANX3、GPR20、LY6K、OR51E2、TARP、WTl、NY-ESO-1、LAGE-la、MAGE-Al、豆莢蛋白、HPV E6、E7、MAGE Al、ETV6-AML、精子蛋白17、XAGE1、Tie 2、MAD-CT-1、MAD-CT-2、Fos相關抗原1、p53、p53突變體、普洛斯坦、存活素、端粒酶、PCTA-1/半乳糖凝集素8、MelanA/MARTl、Ras突變體、hTERT、肉瘤易位斷點、ML-IAP、ERG (TMPRSS2 ETS融合基因)、NA17、PAX3、雄激素受體、周期蛋白Bl、MYCN、RhoC、TRP-2、CYP1B1、BORIS、SART3、PAX5、OY-TES1、LCK、AKAP-4、SSX2、RAGE-1、人類端粒酶逆轉錄酶、RU1、RU2、腸道羧基酯酶、mut hsp70-2、CD79a、CD79b、CD72、LAIR1、FCAR、LILRA2、CD300LF、CLEC12A、BST2、EMR2、LY75、GPC3、FCRL5、IGLL1、CD2、CD3ε、CD4、CD5、CD7、該APRIL蛋白之該細胞外部分或其任何組合。在一些態樣中，該TCR靶向AFP、CD19、TRAC、TCRβ、BCMA、CLL-1、CS1、CD38、CD19、TSHR、CD123、CD22、CD30、CD171、CD33、EGFRvIII、GD2、GD3、Tn Ag、PSMA、ROR1、ROR2、GPC1、GPC2、FLT3、FAP、TAG72、CD44v6、CEA、EPCAM、B7H3、KIT、IL-13Ra2、間皮素、IL-l lRa、PSCA、PRSS21、VEGFR2、LewisY、CD24、PDGFR-β、SSEA-4、CD20、葉酸受體α、ERBB2 (Her2/neu)、MUC1、MUC16、EGFR、NCAM、前列腺酶、PAP、ELF2M、艾普瑞林B2、IGF-I受體、CAIX、LMP2、gplOO、bcr-abl、酪胺酸酶、EphA2、岩藻糖基GM1、sLe、GM3、TGS5、HMWMAA、鄰-乙醯基-GD2、葉酸受體β、TEM1/CD248、TEM7R、CLDN6、GPRC5D、CXORF61、CD97、CD179a、ALK、聚唾液酸、PLAC1、GloboH、NY-BR-1、UPK2、HAVCR1、ADRB3、PANX3、GPR20、LY6K、OR51E2、TARP、WTl、NY-ESO-1、LAGE-la、MAGE-Al、豆莢蛋白、HPV E6、E7、MAGE Al、ETV6-AML、精子蛋白17、XAGE1、Tie 2、MAD-CT-1、MAD-CT-2、Fos相關抗原1、p53、p53突變體、普洛斯坦、存活素、端粒酶、PCTA-1/半乳糖凝集素8、MelanA/MARTl、Ras突變體、hTERT、肉瘤易位斷點、ML-IAP、ERG (TMPRSS2 ETS融合基因)、NA17、PAX3、雄激素受體、周期蛋白Bl、MYCN、RhoC、TRP-2、CYP1B1、BORIS、SART3、PAX5、OY-TES1、LCK、AKAP-4、SSX2、RAGE-1、人類端粒酶逆轉錄酶、RU1、RU2、腸道羧基酯酶、mut hsp70-2、CD79a、CD79b、CD72、LAIR1、FCAR、LILRA2、CD300LF、CLEC12A、BST2、EMR2、LY75、GPC3、FCRL5、IGLL1、CD2、CD3ε、CD4、CD5、CD7、APRIL蛋白之細胞外部分及其任何組合。 II.C.  核酸分子In some aspects, the bispecific antibody comprises (i) an anti-CD161 antibody or an antigen-binding portion thereof disclosed herein and (ii) an antibody or an antigen-binding portion thereof that specifically binds to a tumor antigen. In some aspects, the tumor antigen is selected from CD19, TRAC, TCRβ, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD70, CD171, CD33, EGFRvIII, GD2, GD3, Tn Ag, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, mesothelin, IL-1 lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-β, SSEA-4, CD20, folate receptor α, ERBB2 (Her2/neu), MUC1, MUC16, EGFR, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor β, TEM1/CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WTl, NY-ESO-1, LAGE-la, MAGE-Al, legumain, HPV E6, E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, protan, survivin, telomerase, PCTA-1/galectin 8, MelanA/MART1, Ras mutant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin B1, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mut The extracellular portion of hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, CD2, CD3ε, CD4, CD5, CD7, APRIL protein, or any combination thereof. In some aspects, the TCR targets AFP, CD19, TRAC, TCRβ, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD171, CD33, EGFRvIII, GD2, GD3, Tn Ag, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, mesothelin, IL-1 lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-β, SSEA-4, CD20, folate receptor α, ERBB2 (Her2/neu), MUC1, MUC16, EGFR, NCAM, prostate enzyme, PAP, ELF2M, aprelin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor β, TEM1/CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-Al, soy protein, HPV E6, E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostatin, survivin, telomerase, PCTA-1/galectin 8, MelanA/MART1, Ras mutant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin B1, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mut The extracellular portion of hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, CD2, CD3ε, CD4, CD5, CD7, APRIL protein, and any combination thereof. In some aspects, the bispecific antibody comprises (i) an anti-CD161 antibody disclosed herein, or an antigen-binding portion thereof, and (ii) an antibody or an antigen-binding portion thereof that specifically binds to a tumor antigen selected from the group consisting of CD19, TRAC, TCRβ, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD70, CD171, CD33, EGFRvIII, GD2, GD3, Tn Ag, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, mesothelin, IL-1 lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-β, SSEA-4, CD20, folate receptor α, ERBB2 (Her2/neu), MUC1, MUC16, EGFR, NCAM, prostate enzyme, PAP, ELF2M, aprelin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor β, TEM1/CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-Al, soy protein, HPV E6, E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostatin, survivin, telomerase, PCTA-1/galectin 8, MelanA/MART1, Ras mutant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin B1, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, CD2, CD3ε, CD4, CD5, CD7, the extracellular portion of the APRIL protein, or any combination thereof. In some aspects, the TCR targets AFP, CD19, TRAC, TCRβ, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD171, CD33, EGFRvIII, GD2, GD3, Tn Ag, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, mesothelin, IL-1 lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-β, SSEA-4, CD20, folate receptor α, ERBB2 (Her2/neu), MUC1, MUC16, EGFR, NCAM, prostate enzyme, PAP, ELF2M, aprelin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor β, TEM1/CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-Al, soy protein, HPV E6, E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostatin, survivin, telomerase, PCTA-1/galectin 8, MelanA/MART1, Ras mutant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin B1, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mut Extracellular portions of hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, CD2, CD3ε, CD4, CD5, CD7, APRIL proteins, and any combination thereof.II.C. Nucleic acid molecules

本文所述的另一個態樣係關於編碼本文所述的抗CD161抗體之核酸分子。核酸可存在於整個細胞、細胞溶解產物中、或以部分純化或實質上純淨之形式存在。核酸在藉由標準技術(包括鹼/SDS處理、CsCl顯帶、管柱層析、限制酶、瓊脂糖凝膠電泳及此項技術中熟知的其他技術)自其他細胞組分或其他污染物(例如其他細胞核酸(例如其他染色體DNA，例如天然連接至分離的DNA之染色體DNA)或蛋白質)純化時「經分離」或「致使變得實質上純」。參見，F. Ausubel等人編(1987) Current Protocols in Molecular Biology, Greene Publishing及Wiley Interscience, New York。本文所述的核酸可為例如DNA或RNA且可含有或可不含有內含子序列。在一些態樣中，該核酸係cDNA分子。Another aspect described herein is about the nucleic acid molecule of the anti-CD161 antibody described herein. Nucleic acid can be present in whole cell, cell lysis product or in partially purified or substantially pure form. Nucleic acid is "separated" or "caused to become substantially pure" when purified from other cell components or other contaminants (such as other cell nucleic acids (such as other chromosome DNA, such as chromosome DNA naturally connected to the separated DNA) or protein) by standard techniques (including alkali/SDS treatment, CsCl banding, column chromatography, restriction enzymes, agarose gel electrophoresis and other techniques known in this technology). Refer to, F. Ausubel et al. (1987) Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York. Nucleic acid described herein can be, for example, DNA or RNA and may or may not contain intron sequences. In some aspects, the nucleic acid is a cDNA molecule.

本文所述的核酸可使用標準分子生物學技術來獲得。對於藉由雜交瘤(例如自攜載如下文進一步描述的人類免疫球蛋白基因之轉殖基因小鼠製備的雜交瘤)表現之抗體，編碼藉由該雜交瘤製造的抗體之重鏈及輕鏈之cDNA可藉由標準PCR擴增或cDNA選殖技術獲得。對於自免疫球蛋白基因庫(例如使用噬菌體展示技術)獲得的抗體，可自該庫移除編碼該抗體之核酸。The nucleic acids described herein can be obtained using standard molecular biology techniques. For antibodies expressed by hybridomas (e.g., hybridomas prepared from transgenic mice carrying human immunoglobulin genes as described further below), cDNAs encoding the heavy and light chains of the antibodies produced by the hybridomas can be obtained by standard PCR amplification or cDNA cloning techniques. For antibodies obtained from immunoglobulin gene libraries (e.g., using phage display technology), nucleic acids encoding the antibodies can be removed from the library.

本文所述的一些核酸分子係彼等編碼本文揭示的抗CD161抗體之VH及VL序列者。Some of the nucleic acid molecules described herein are those encoding the VH and VL sequences of the anti-CD161 antibodies disclosed herein.

本發明之核酸分子可經修飾以刪除特定序列(例如限制酶識別序列)、或最佳化密碼子。The nucleic acid molecules of the present invention can be modified to delete specific sequences (such as restriction enzyme recognition sequences) or to optimize codons.

一種用於製造本文揭示的抗CD161抗體之方法可包含在包含編碼重鏈及輕鏈之核苷酸序列之細胞系中用信號肽表現該等重鏈及輕鏈。本文涵蓋包含此等核苷酸序列之宿主細胞。A method for producing an anti-CD161 antibody disclosed herein may include expressing the heavy and light chains using a signal peptide in a cell line comprising nucleotide sequences encoding the heavy and light chains. Host cells comprising these nucleotide sequences are contemplated herein.

一旦獲得編碼VH及VL區段之DNA片段，此等DNA片段可進一步藉由標準重組DNA技術操縱，例如以將可變區基因轉化至全長抗體鏈基因、Fab片段基因或scFv基因。在此等操縱中，VL-或VH編碼DNA片段可操作地連接至編碼另一蛋白質之另一DNA片段，諸如抗體恆定區或可撓連接子。術語「可操作地連接」如此上下文中所使用意欲意指兩個DNA片段經接合使得由兩個DNA片段編碼之胺基酸序列保持在同框。Once the DNA fragments encoding the VH and VL segments are obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example, to convert the variable region genes into full-length antibody chain genes, Fab fragment genes, or scFv genes. In these manipulations, the VL- or VH-encoding DNA fragment is operably linked to another DNA fragment encoding another protein, such as an antibody constant region or a flexible linker. The term "operably linked" as used in this context is intended to mean that the two DNA fragments are joined so that the amino acid sequences encoded by the two DNA fragments remain in the same frame.

編碼VH區之分離的DNA可藉由將VH編碼DNA可操作地連接至編碼重鏈恆定區(鉸鏈、CH1、CH2及/或CH3)之另一DNA分子而轉化為全長重鏈基因。人類重鏈恆定區基因之序列係此項技術中已知的(參見，例如，Kabat, E. A.等人(1991) Sequences of Proteins of Immunological Interest, 第五版, U.S. Department of Health and Human Services, NIH出版91-3242號)且涵蓋此等區域之DNA片段可藉由標準PCR擴增來獲得。重鏈恆定區可係IgG1、IgG2、IgG3、IgG4、IgA、IgE、IgM或IgD恆定區，例如IgG1區。對於Fab片段重鏈基因，VH編碼DNA可可操作地連接至僅編碼重鏈CH1恆定區之另一DNA分子。The isolated DNA encoding the VH region can be converted to a full-length heavy chain gene by operably linking the VH encoding DNA to another DNA molecule encoding a heavy chain constant region (hinge, CH1, CH2 and/or CH3). The sequences of human heavy chain constant region genes are known in the art (see, e.g., Kabat, E.A. et al. (1991) Sequences of Proteins of Immunological Interest, 5th edition, U.S. Department of Health and Human Services, NIH publication No. 91-3242) and DNA fragments covering these regions can be obtained by standard PCR amplification. The heavy chain constant region can be an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region, e.g., an IgG1 region. For Fab fragment heavy chain genes, the VH encoding DNA can be operably linked to another DNA molecule encoding only the heavy chain CH1 constant region.

編碼VL區之分離的DNA可藉由將VL編碼DNA可操作地連接至編碼輕鏈恆定區CL之另一DNA分子轉化為全長輕鏈基因(以及Fab輕鏈基因)。人類輕鏈恆定區基因之序列係此項技術中已知的(參見，例如，Kabat, E. A.等人(1991) Sequences of Proteins of Immunological Interest, 第五版, U.S. Department of Health and Human Services, NIH出版91-3242號)且涵蓋此等區域之DNA片段可藉由標準PCR擴增來獲得。輕鏈恆定區可係κ或λ恆定區。The isolated DNA encoding the VL region can be converted into a full-length light chain gene (as well as a Fab light chain gene) by operably linking the VL encoding DNA to another DNA molecule encoding the light chain constant region CL. The sequences of human light chain constant region genes are known in the art (see, e.g., Kabat, E.A. et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242) and DNA fragments covering these regions can be obtained by standard PCR amplification. The light chain constant region can be a kappa or lambda constant region.

為了產生scFv基因，VH-及VL編碼DNA片段可操作地連接至編碼可撓連接子(例如編碼胺基酸序列(Gly₄-Ser)₃)之另一片段，使得VH及VL序列可經表現為連續單鏈蛋白，其中該等VL及VH區經該可撓連接子接合。To generate scFv genes, VH- and VL-encoding DNA fragments are operably linked to another fragment encoding a flexible linker (e.g., encoding the amino acid sequence (_Gly4 -Ser)₃ ) so that the VH and VL sequences can be expressed as a continuous single chain protein in which the VL and VH regions are joined by the flexible linker.

本發明之一些態樣係關於一種載體或一組載體，其包含本文揭示的核酸分子。在一些態樣中，該載體係病毒載體。在一些態樣中，該載體係病毒顆粒或病毒。在一些態樣中，該載體係哺乳動物載體。在一些態樣中，該載體係細菌載體。Some aspects of the invention relate to a vector or a set of vectors comprising a nucleic acid molecule disclosed herein. In some aspects, the vector is a viral vector. In some aspects, the vector is a viral particle or a virus. In some aspects, the vector is a mammalian vector. In some aspects, the vector is a bacterial vector.

在某些態樣中，該載體係逆轉錄病毒載體。在一些態樣中，該載體選自腺病毒載體、慢病毒、仙台病毒(Sendai virus)、桿狀病毒載體、埃巴病毒載體(Epstein Barr viral vector)、乳頭多瘤空泡病毒載體(papovaviral vector)、牛痘病毒載體、單純疱疹病毒載體及腺相關病毒(AAV)載體。在特定態樣中，該載體係AAV載體。在一些態樣中，該載體係慢病毒。在特定態樣中，該載體係AAV載體。在一些態樣中，該載體係仙台病毒。在一些態樣中，該載體係雜交載體。可用於本發明中之雜交載體之實例可見於Huang及Kamihira,Biotechnol. Adv. 31(2):208-23 (2103)，其全文以引用之方式併入本文中。In some aspects, the vector is a retroviral vector. In some aspects, the vector is selected from adenoviral vectors, lentivirus, Sendai virus, rod-shaped virus vectors, Epstein Barr viral vectors, papovaviral vectors, vaccinia virus vectors, herpes simplex virus vectors, and adeno-associated virus (AAV) vectors. In certain aspects, the vector is an AAV vector. In certain aspects, the vector is a lentivirus. In certain aspects, the vector is an AAV vector. In certain aspects, the vector is a Sendai virus. In certain aspects, the vector is a hybrid vector. Examples of hybridization vectors that can be used in the present invention can be found in Huang and Kamihira,Biotechnol. Adv. 31(2) :208-23 (2103), which is incorporated herein by reference in its entirety.

本發明之一些態樣係關於包含本文揭示之核酸分子、一組核酸分子、載體或一組載體之宿主細胞。在一些態樣中，該宿主細胞係哺乳動物細胞。在一些態樣中，該宿主細胞係活體外細胞。 II.D.  醫藥組合物Some aspects of the invention relate to a host cell comprising a nucleic acid molecule, a set of nucleic acid molecules, a vector, or a set of vectors disclosed herein. In some aspects, the host cell is a mammalian cell. In some aspects, the host cell is an ex vivo cell.II.D. Pharmaceutical Compositions

進一步提供包含本文揭示之抗CD161抗體、核酸分子、載體或宿主細胞及一或多種醫藥上可接受之載劑之組合物(例如醫藥組合物)。Further provided are compositions (eg, pharmaceutical compositions) comprising the anti-CD161 antibodies, nucleic acid molecules, vectors or host cells disclosed herein and one or more pharmaceutically acceptable carriers.

在一些態樣中，該組合物進一步包含增積劑。增積劑可選自由NaCl、甘露醇、甘胺酸、丙胺酸及其任何組合組成之群。在其他態樣中，該組合物包含穩定劑。該穩定劑可選自由蔗糖、海藻糖、棉子糖、精胺酸；或其任何組合組成之群。在其他態樣中，該組合物包含表面活性劑。在一些態樣中，該表面活性劑選自聚山梨醇酯80 (Ps80)、聚山梨醇酯20 (PS20)及其任何組合。在其他態樣中，該組合物進一步包含螯合劑。在一些態樣中，該螯合劑選自二乙三胺五乙酸(DTPA)、乙二胺四乙酸、氮基三乙酸及其任何組合。在一些態樣中，該組合物進一步包含NaCl、甘露醇、噴替酸(pentetic acid) (DTPA)、蔗糖、PS80或其任何組合。In some embodiments, the composition further comprises an accumulator. The accumulator may be selected from the group consisting of NaCl, mannitol, glycine, alanine, and any combination thereof. In other embodiments, the composition comprises a stabilizer. The stabilizer may be selected from the group consisting of sucrose, trehalose, raffinose, arginine; or any combination thereof. In other embodiments, the composition comprises a surfactant. In some embodiments, the surfactant is selected from polysorbate 80 (Ps80), polysorbate 20 (PS20), and any combination thereof. In other embodiments, the composition further comprises a chelating agent. In some embodiments, the chelating agent is selected from diethylenetriaminepentaacetic acid (DTPA), ethylenediaminetetraacetic acid, nitrilotriacetic acid, and any combination thereof. In some aspects, the composition further comprises NaCl, mannitol, pentetic acid (DTPA), sucrose, PS80, or any combination thereof.

如本文所用，「醫藥上可接受之載劑」包括任何及所有的生理上相容之溶劑、分散介質、塗料、抗細菌劑及抗真菌劑、等滲劑及吸收延遲劑及類似者。在一些態樣中，該載劑適合於靜脈內、肌肉內、皮下、非經腸、脊髓或表皮投與(例如藉由注射或輸注)。取決於投與途徑，該活性化合物(亦即抗體、免疫結合物或雙特異性分子)可塗佈於材料中以保護該化合物免受酸及可使該化合物去活化之其他天然條件的作用的影響。As used herein, "pharmaceutically acceptable carriers" include any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. In some aspects, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound (i.e., antibody, immunoconjugate or bispecific molecule) can be coated in a material to protect the compound from the effects of acids and other natural conditions that can deactivate the compound.

本文所述的醫藥化合物可包括一或多種醫藥上可接受之鹽。「醫藥上可接受之鹽」係指保留親本化合物之所需生物活性且不賦予任何非所欲毒理效應之鹽(參見，例如，Berge, S.M.等人(1977)J. Pharm. Sci.66: 1-19)。此類鹽之實例包括酸加成鹽及鹼加成鹽。酸加成鹽包括彼等衍生自非毒性無機酸(諸如鹽酸、硝酸、磷酸、硫酸、氫溴酸、氫碘酸、亞磷酸及類似者)之鹽以及彼等衍生自非毒性有機酸(諸如脂族單羧酸及二羧酸、苯基取代之烷酸、羥基烷酸、芳族酸、脂族及芳族磺酸及類似者)之鹽。鹼加成鹽包括彼等衍生自鹼土金屬(諸如鈉、鉀、鎂、鈣及類似者)之鹽以及彼等衍生自非毒性有機胺(諸如Ν,Ν'-二苄基乙二胺、N-甲基葡糖胺、氯普魯卡因、膽鹼、二乙醇胺、乙二胺、普魯卡因(procaine)及類似者)之鹽。The pharmaceutical compounds described herein may include one or more pharmaceutically acceptable salts. A "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the parent compound and does not impart any undesirable toxicological effects (see, e.g., Berge, SM et al. (1977)J. Pharm. Sci. 66: 1-19). Examples of such salts include acid addition salts and base addition salts. Acid addition salts include those derived from non-toxic inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphorous acid, and the like, and those derived from non-toxic organic acids such as aliphatic monocarboxylic acids and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and the like. Base addition salts include those derived from alkaline earth metals such as sodium, potassium, magnesium, calcium and the like and those derived from non-toxic organic amines such as N,N'-dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine and the like.

本文描述的醫藥組合物亦可包括醫藥上可接受之抗氧化劑。醫藥上可接受之抗氧化劑之實例包括：(1)水溶性抗氧化劑，諸如抗壞血酸、半胱胺酸鹽酸鹽、硫酸氫鈉、偏亞硫酸氫鈉、亞硫酸鈉及類似者；(2)油溶性抗氧化劑，諸如抗壞血酸棕櫚酸酯、丁基化羥基苯甲醚(BHA)、丁基化羥基甲苯(BHT)、卵磷脂、没食子酸丙酯、α-生育酚及類似者；及(3)金屬螯合劑，諸如檸檬酸、乙二胺四乙酸(EDTA)、山梨糖醇、酒石酸、磷酸及類似者。The pharmaceutical compositions described herein may also include a pharmaceutically acceptable antioxidant. Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants such as ascorbic acid palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, and the like; and (3) metal chelators such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

可用於本文描述的醫藥組合物中之適宜水性及非水性載劑之實例包括水、乙醇、多元醇(諸如甘油、丙二醇、聚乙二醇及類似者)、及其適宜混合物、植物油(諸如橄欖油)、及可注射有機酯(諸如油酸乙酯)。可例如藉由使用塗覆材料(諸如卵磷脂)、就分散液而言藉由維持所需粒度、及藉由使用表面活性劑來維持恰當的流動性。Examples of suitable aqueous and nonaqueous carriers that can be used in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters (such as ethyl oleate). Proper fluidity can be maintained, for example, by the use of coating materials (such as lecithin), by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

此等組合物亦可含有佐劑，諸如防腐劑、潤濕劑、乳化劑及分散劑。可藉由滅菌程序(同上)、及藉由包含各種抗細菌劑及抗真菌劑(例如對羥基苯甲酸酯、氯丁醇、苯酚山梨酸及類似者)來確保防止微生物之存在。亦可期望包括等滲劑(諸如糖、氯化鈉及類似者)至該等組合物中。此外，該可注射醫藥形式之延長吸收可藉由包含延遲吸收之試劑(諸如單硬脂酸鋁及明膠)來實現。These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers and dispersants. Prevention of the presence of microorganisms can be ensured by sterilization procedures (same as above), and by the inclusion of various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol sorbic acid and the like). It may also be desirable to include isotonic agents (e.g., sugars, sodium chloride and the like) into these compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be achieved by the inclusion of agents that delay absorption (e.g., aluminum monostearate and gelatin).

醫藥上可接受之載劑包括無菌水性溶液或用於即時製備無菌可注射溶液或分散液之分散液及無菌粉末。此類介質及藥劑用於醫藥活性物質之用途係此項技術中已知的。除非任何習知介質或藥劑與活性化合物不相容，否則考慮將其用於本文描述的醫藥組合物中。醫藥組合物可包含防腐劑且可不含防腐劑。可將補充活性化合物併入至組合物中。Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is known in the art. Unless any known media or agents are incompatible with the active compound, they are contemplated for use in the pharmaceutical compositions described herein. The pharmaceutical compositions may contain preservatives and may be preservative-free. Supplementary active compounds may be incorporated into the compositions.

在製造及儲存之條件下，治療性組合物通常必須是無菌且穩定。該組合物可經調配為適合於高藥物濃度之溶液、微乳液、脂質體或其他有序結構。該載劑可係溶劑或分散介質，其含有例如水、乙醇、多元醇(例如甘油、丙二醇及液體聚乙二醇及類似者)及其適宜混合物。可例如藉由使用塗料(諸如卵磷脂)、就分散液而言藉由維持所需粒度、及藉由使用表面活性劑來維持恰當的流動性。在許多情況下，該等組合物可在該組合物中包括等滲劑，例如糖、多元醇(諸如甘露醇)、山梨糖醇或氯化鈉。可注射組合物的延長吸收可藉由在該組合物中包含延遲吸收的試劑(例如單硬脂酸鹽及明膠)來實現。The therapeutic composition must generally be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome or other ordered structure suitable for high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyols (such as glycerol, propylene glycol and liquid polyethylene glycol and the like) and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of coatings (such as lecithin), by maintaining the desired particle size in the case of dispersions, and by the use of surfactants. In many cases, the compositions may include isotonic agents in the composition, such as sugars, polyols (such as mannitol), sorbitol or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, monostearate salts and gelatin.

無菌可注射溶液可藉由將該活性化合物以所需量視需要與以上所列舉的成分中之一者或以上所列舉的成分之組合併入於適宜溶劑中接著進行滅菌微過濾來製備。一般而言，分散液藉由將活性化合物併入至無菌媒劑中來製備，該無菌媒劑含有基礎分散介質及所需的其他選自彼等本文所列舉者之成分。就用於製備無菌可注射溶液之無菌粉末而言，一些製備方法係自其先前經無菌過濾之溶液產生活性成分加上任一另外所需成分之粉末之真空乾燥及冷凍乾燥(凍乾)。Sterile injectable solutions can be prepared by incorporating the active compound in the desired amount in an appropriate solvent, optionally with one of the ingredients listed above or a combination of the ingredients listed above, followed by sterilization microfiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and other ingredients selected from those listed herein as desired. For sterile powders for the preparation of sterile injectable solutions, some preparation methods are vacuum drying and freeze drying (lyophilization) of a powder of the active ingredient plus any additional desired ingredients from a previously sterile filtered solution thereof.

可與載劑材料組合以產生單一劑型之活性成分之量將取決於所治療的個體及特定投與模式而變化。可與載劑材料混合以產生單一劑型之活性成分之量將一般係該組合物之產生治療效應之量。一般而言，在100%中，此量將在約0.01%至約99%之活性成分、約0.1%至約70%、或約1%至約30%之活性成分與醫藥上可接受之載劑之組合之範圍內。The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the individual being treated and the particular mode of administration. The amount of active ingredient that can be mixed with a carrier material to produce a single dosage form will generally be that amount of the composition that produces a therapeutic effect. Generally speaking, out of 100%, this amount will be in the range of about 0.01% to about 99% of the active ingredient, about 0.1% to about 70%, or about 1% to about 30% of the active ingredient in combination with a pharmaceutically acceptable carrier.

本文描述的組合物可使用此項技術中已知的多種方法中之一者或多者經由一或多種投與途徑來投與。如熟練技術者所瞭解，該投與途徑及/或模式將取決於所需結果而變化。本文描述的抗CD161抗體之投與途徑包括靜脈內、肌肉內、皮下、腹膜內、脊髓或其他非經腸投與途徑，例如藉由注射或輸注。The compositions described herein may be administered via one or more routes of administration using one or more of a variety of methods known in the art. As will be appreciated by the skilled artisan, the route and/or mode of administration will vary depending on the desired outcome. Routes of administration of the anti-CD161 antibodies described herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other non-parenteral routes of administration, such as by injection or infusion.

活性化合物可用將保護化合物免於快速釋放之載劑製備，諸如控制釋放型調配物，包括植入物、經皮貼劑及微膠囊化遞送系統。可使用生物可降解、生物相容性聚合物，諸如乙烯乙酸乙烯酯、聚酸酐、聚乙醇酸、膠原蛋白、聚原酸酯及聚乳酸。用於製備此類調配物之許多方法為專利化或為熟習此項技術者一般已知。參見，例如，Sustained and Controlled Release Drug Delivery Systems, J.R.Robinson編, Marcel Dekker, Inc., New York, 1978。 III.   本發明方法 III.A. 治療方法The active compound may be prepared with a carrier that will protect the compound from rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid may be used. Many methods for preparing such formulations are patented or generally known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.III.   Methods of the Invention III.A. Methods of Treatment

本發明之一些態樣係關於治療個體之疾病或病症之方法，其包括對該個體投與本文揭示的抗CD161抗體、編碼該抗CD161抗體之多核苷酸、包含該多核苷酸之載體、包含該多核苷酸之宿主細胞、或其任何組合。在一些態樣中，該疾病或病症包含癌症。Some aspects of the present invention are about the method for treating individual disease or illness, which includes administering anti-CD161 antibodies disclosed herein, polynucleotides encoding these anti-CD161 antibodies, vectors comprising these polynucleotides, host cells comprising these polynucleotides, or any combination thereof. In some aspects, these diseases or illnesses comprise cancer.

本發明之一些態樣係關於一種治療有需要個體之癌症之方法，其包括對該個體投與有效劑量之本文揭示的組合物(例如抗體、多核苷酸、載體、宿主細胞或醫藥組合物)。在其他態樣中，本發明係關於一種殺死有需要個體之腫瘤細胞之方法，其包括對該個體投與有效劑量之本文揭示的組合物。在其他態樣中，本發明係關於一種減小有需要個體之腫瘤之尺寸之方法，其包括對該個體投與有效劑量之本文揭示的組合物。在其他態樣中，本發明係關於抑制有需要個體之腫瘤之轉移，其包括對該個體投與有效劑量之本文揭示的組合物。在一些態樣中，該個體係人類。Some aspects of the present invention relate to a method of treating cancer in an individual in need thereof, comprising administering to the individual an effective dose of a composition disclosed herein (e.g., an antibody, polynucleotide, vector, host cell, or pharmaceutical composition). In other aspects, the present invention relates to a method of killing tumor cells in an individual in need thereof, comprising administering to the individual an effective dose of a composition disclosed herein. In other aspects, the present invention relates to a method of reducing the size of a tumor in an individual in need thereof, comprising administering to the individual an effective dose of a composition disclosed herein. In other aspects, the present invention relates to inhibiting the metastasis of a tumor in an individual in need thereof, comprising administering to the individual an effective dose of a composition disclosed herein. In some aspects, the individual is a human.

本發明之一些態樣係關於誘導有需要個體中之免疫反應之方法，其包括對該個體投與本文揭示之抗CD161抗體、編碼該抗CD161抗體之多核苷酸、包含該多核苷酸之載體、包含該多核苷酸之宿主細胞或其任何組合。在一些態樣中，該個體罹患癌症。Some aspects of the present invention are about inducing the method for the immune response in the individual in need, which includes administering the anti-CD161 antibody disclosed herein, the polynucleotides encoding the anti-CD161 antibody, the vector comprising the polynucleotides, the host cell comprising the polynucleotides or any combination thereof to the individual. In some aspects, the individual suffers from cancer.

本發明之一些態樣係關於一種活化免疫細胞之方法，其包括使該免疫細胞與本文揭示之抗CD161抗體、編碼該抗CD161抗體之多核苷酸、包含該多核苷酸之載體、包含該多核苷酸之宿主細胞、或其任何組合接觸。在一些態樣中，該免疫細胞係活體外接觸。在一些態樣中，該免疫細胞係離體接觸。在一些態樣中，該免疫細胞存在於人類個體中，亦即，該免疫細胞係活體內接觸。在一些態樣中，該個體罹患癌症。Some aspects of the present invention are related to a method of activating immune cells, which includes contacting the immune cells with anti-CD161 antibodies disclosed herein, polynucleotides encoding the anti-CD161 antibodies, vectors comprising the polynucleotides, host cells comprising the polynucleotides, or any combination thereof. In some aspects, the immune cells are contacted in vitro. In some aspects, the immune cells are contacted in vitro. In some aspects, the immune cells are present in human individuals, that is, the immune cells are contacted in vivo. In some aspects, the individual suffers from cancer.

在一些態樣中，該癌症包含實體腫瘤。在一些態樣中，該癌症包含惡性血液病。在一些態樣中，該癌症為局部晚期。在一些態樣中，該癌症為轉移。在一些態樣中，該腫瘤為復發性。在一些態樣中，該腫瘤為難治性。在一些態樣中，該腫瘤在一或多種先前的治療該腫瘤之療法後為復發性及/或難治性。在一些態樣中，該一或多種先前療法包含標準照護療法。在一些態樣中，該一或多種先前療法包含化學療法。在一些態樣中，該一或多種先前療法包含免疫療法。在一些態樣中，該一或多種先前療法包含外科手術。在一些態樣中，該一或多種先前療法包含放射療法。In some aspects, the cancer comprises a solid tumor. In some aspects, the cancer comprises a hematological malignancy. In some aspects, the cancer is locally advanced. In some aspects, the cancer is metastatic. In some aspects, the tumor is recurrent. In some aspects, the tumor is refractory. In some aspects, the tumor is recurrent and/or refractory after one or more prior treatments for the tumor. In some aspects, the one or more prior treatments comprise standard of care. In some aspects, the one or more prior treatments comprise chemotherapy. In some aspects, the one or more prior treatments comprise immunotherapy. In some aspects, the one or more prior treatments comprise surgery. In some aspects, the one or more prior treatments include radiation therapy.

在一些態樣中，該癌症選自聽神經瘤、急性淋巴球性白血病、急性髓細胞性白血病、腺癌、及泌尿系統之癌症、及上皮細胞癌、血管肉瘤、星形細胞瘤、基底細胞癌、膽管癌、膽道癌、膀胱癌、骨癌、腦癌、腦幹神經膠質瘤、乳癌、支氣管癌、伯奇氏淋巴瘤及邊緣區B細胞淋巴瘤、腎上腺癌、肛門區癌、消化系統癌、內分泌系統癌、食道癌、副甲狀腺癌、陰莖癌、呼吸系统癌、小腸癌、輸尿管癌、尿道癌、子宮頸癌、子宮內膜癌、輸卵管癌、腎盂癌、陰道癌、外陰癌、中樞神經系統(CNS)癌症、子宮頸癌、軟骨肉瘤、脊索瘤、絨毛膜癌、慢性白血病、慢性淋巴球性白血病、慢性髓細胞性(粒細胞性)白血病、結腸癌、結腸肉瘤、結腸直腸癌、結締組織癌、顱咽管瘤、囊腺癌、胚胎癌、子宮內膜癌、內皮肉瘤、環境誘導型癌症(包括彼等由石棉誘導者)、室管膜瘤、表皮樣癌、上皮樣癌、食道癌、食道上皮細胞癌、尤因氏肉瘤、眼癌、纖維肉瘤、胃癌(gastric cancer)、胃腸癌、生殖細胞腫瘤、神經膠質母細胞瘤(例如多形性神經膠質母細胞瘤)、神經膠質瘤、頭頸癌、重鏈病、血管母細胞瘤、肝癌瘤、霍奇金氏病、上皮內贅瘤、卡波西氏肉瘤、腎癌(例如腎細胞癌(RCC))、喉頭癌、平滑肌肉瘤、白血病、脂肪肉瘤、肝癌、肺癌(lung cancer) (小細胞、大細胞)、肺癌(lung carcinoma)、淋巴管內皮肉瘤、淋巴管肉瘤、被套細胞淋巴瘤、髓質癌、神經管胚細胞瘤、黑色素瘤、腦膜瘤、間皮瘤、多發性骨髓瘤、骨髓母細胞前髓細胞骨髓單核球性單核細胞紅血球性白血病、黏液肉瘤、鼻咽癌、中樞神經系統(CNS)贅瘤、神經母細胞瘤、非霍奇金氏病、非小細胞肺癌(non-small cell lung cancer；NSCLC)、非小細胞肺上皮細胞癌(non-small cell lung carcinoma)、寡樹突神經膠質瘤、口腔癌(例如唇、舌、口及咽)、成骨肉瘤、骨肉瘤、卵巢癌、胰臟癌、乳突腺癌、乳突癌、小兒肉瘤、松果腺瘤、垂體腺瘤、真性紅血球增多症、淋巴瘤、原發性CNS淋巴瘤、前列腺癌(例如激素難治性前列腺腺癌)、直腸癌、腎癌(例如透明細胞癌)、視網膜母細胞瘤、橫紋肌肉瘤、肉瘤、軟組織肉瘤、皮脂腺癌、精細胞瘤、鼻腔鼻竇自然殺手、皮膚癌、小細胞肺癌(SCLC)、兒童實體腫瘤、脊髓軸瘤、鱗狀細胞癌(squamous cell cancer)、鱗狀細胞癌(squamous cell carcinoma)、胃癌(stomach cancer)、汗腺癌、滑膜瘤、睾丸癌、甲狀腺癌、腫瘤血管生成、子宫癌、病毒相關癌症或病毒起源之癌症(例如人類乳頭瘤病毒(HPV相關或來源腫瘤))、華氏巨球蛋白血症及威爾姆氏腫瘤；及該等癌症之任何組合。In some embodiments, the cancer is selected from acoustic neuroma, acute lymphocytic leukemia, acute myeloid leukemia, adenocarcinoma, and cancer of the urinary system, and epithelial cell carcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct carcinoma, bile duct cancer, bladder cancer, bone cancer, brain cancer, brain stem neuroglioma, breast cancer, bronchial cancer, Birch's lymphoma and marginal zone B cell lymphoma, adrenal cancer, anal cancer, digestive system cancer, endocrine system cancer, esophageal cancer, parathyroid cancer, penile cancer, respiratory system cancer, small intestine cancer, ureteral cancer, urethral cancer, cervical cancer, endometrial cancer, fallopian tube cancer, cancer, renal pelvic cancer, vaginal cancer, vulvar cancer, central nervous system (CNS) cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, colon cancer, colon sarcoma, colorectal cancer, connective tissue cancer, cranio-pharyngioma, cystadenocarcinoma, embryonal carcinoma, endometrial cancer, endothelial sarcoma, environmentally induced cancers (including those induced by asbestos), ependymoma, epidermoid carcinoma, epithelioid carcinoma, esophageal cancer, esophageal epithelial cell carcinoma, Ewing's sarcoma, eye cancer, fibrosarcoma, gastric cancer (gastric cancer), cancer), gastrointestinal cancer, germ cell tumor, neuroglioblastoma (e.g., multiforme neuroglioblastoma), neuroglioblastoma, head and neck cancer, heavy chain disease, hemangioblastoma, hepatocarcinoma, Hodgkin's disease, intraepithelial neoplasm, Kaposi's sarcoma, kidney cancer (e.g., renal cell carcinoma (RCC)), laryngeal cancer, leiomyosarcoma, leukemia, liposarcoma, liver cancer, lung cancer (small cell, large cell), lung cancer (lung carcinoma), lymphangioendothelioma, lymphangiosarcoma, mantle cell lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloblastic promyelocytic myelomonocytic monocytic erythroblastic leukemia, myxosarcoma, nasopharyngeal carcinoma, central nervous system (CNS) neoplasms, neuroblastoma, non-Hodgkin's disease, non-small cell lung cancer (NSCLC), non-small cell lung epithelial carcinoma (NSCLC), carcinoma), oligodendrocyte neuroglia, oral cancer (e.g., lip, tongue, mouth, and pharynx), osteogenic sarcoma, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pediatric sarcoma, pineal adenoma, pituitary adenoma, polycythemia vera, lymphoma, primary CNS lymphoma, prostate cancer (e.g., hormone-refractory prostate adenocarcinoma), rectal cancer, kidney cancer (e.g., clear cell carcinoma), retinoblastoma, rhabdomyosarcoma, sarcoma, Soft tissue sarcoma, sebaceous gland carcinoma, spermatoma, nasal sinus natural killer, skin cancer, small cell lung cancer (SCLC), solid tumors in children, spinal cord axonal tumors, squamous cell cancer, squamous cell carcinoma carcinoma), stomach cancer, sweat gland cancer, synovialoma, testicular cancer, thyroid cancer, tumor angiogenesis, uterine cancer, virus-related cancers or cancers of viral origin (such as human papillomavirus (HPV-related or derived tumors)), Waldenstrom's macroglobulinemia and Wilms' tumor; and any combination of these cancers.

在一些態樣中，該個體罹患傳染病，亦即，在一些態樣中，該疾病或病症包含傳染病。在一些態樣中，該傳染病選自細菌感染、真菌感染、病毒感染、寄生蟲感染或其任何組合。在一些態樣中，該傳染病包含流行性感冒、皰疹、梨形鞭毛蟲屬、瘧疾、利什曼原蟲或其任何組合之感染。在一些態樣中，該傳染病包含人類免疫缺陷病毒(HIV)、肝炎病毒、皰疹病毒、腺病毒、流行性感冒病毒、黃病毒科、伊科病毒、鼻病毒、柯薩奇病毒、冠狀病毒、呼吸道融合病毒、腮腺炎病毒、輪狀病毒、麻疹病毒、蕁麻疹病毒、小病毒、牛痘病毒、HTLV病毒、登革熱病毒、乳頭瘤病毒、軟疣病毒、脊髓灰質炎病毒、狂犬病病毒、JC病毒、或蟲媒病毒腦炎病毒、或其任何組合之感染。在一些態樣中，該傳染病包含披衣菌、立克次體細菌、分枝桿菌、葡萄球菌、鏈球菌、肺炎球菌、腦膜炎雙球菌、淋球菌、克雷伯氏菌、變形桿菌屬、沙雷氏菌、假單胞菌、軍團菌屬、白喉桿菌、沙門氏菌、桿菌、霍亂、破傷風、肉毒症、炭疽、瘟疫、鉤端螺旋體病、及萊姆氏病細菌、或其任何組合。在一些態樣中，該傳染病包含念珠菌屬、新型隱球菌、麴菌屬、毛黴目屬、申克氏胞絲菌、皮炎芽生菌、巴西副球孢子菌、粗球黴菌、或莢膜組織漿菌、或其任何組合之感染。在一些態樣中，該傳染病包含溶組織內阿米巴、結腸小袋纖毛蟲、福氏耐格里阿米巴原蟲、棘阿米巴科、蘭氏賈第鞭毛蟲、隱孢子蟲、卡式肺囊蟲、間日瘧原蟲、巴貝斯原蟲、布氏錐蟲、克氏錐蟲、杜氏利什曼原蟲、岡地弓漿蟲病、或巴西日圓線蟲、或其任何組合之感染。In some aspects, the individual suffers from an infectious disease, that is, in some aspects, the disease or disorder comprises an infectious disease. In some aspects, the infectious disease is selected from a bacterial infection, a fungal infection, a viral infection, a parasitic infection, or any combination thereof. In some aspects, the infectious disease comprises an infection with influenza, herpes, Giardia, malaria, Leishmania, or any combination thereof. In some aspects, the infectious disease comprises infection with human immunodeficiency virus (HIV), hepatitis virus, herpes virus, adenovirus, influenza virus, Flaviviridae, echovirus, rhinovirus, coxsackie virus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum virus, poliovirus, rabies virus, JC virus, or arbovirus encephalitis virus, or any combination thereof. In some aspects, the infectious disease comprises Chlamydia, Rickettsia, Mycobacterium, Staphylococcus, Streptococcus, Pneumococcus, Meningococcus, Gonorrhea, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Diphtheria, Salmonella, Bacillus, Cholera, Tetanus, Botulism, Anthrax, Plague, Leptospirosis, and Lyme disease bacteria, or any combination thereof. In some aspects, the infectious disease comprises infection with Candida, Cryptococcus neoformans, Aspergillus, Trichoderma, Cytomyces schenckii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, or Histoplasma capsulatum, or any combination thereof. In some aspects, the infectious disease comprises infection with Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoebae, Giardia lamblia, Cryptosporidium, Pneumocystis carinii, Plasmodium vivax, Babesia, Stem Cells, Styrax cruz, Leishmania donovani, Toxoplasma gondii, or Nippleworm brasiliensis, or any combination thereof.

在一些態樣中，該個體罹患自身免疫性疾病。In some aspects, the individual suffers from an autoimmune disease.

本發明組合物可使用任何醫藥上可接受之途徑投與。在一些態樣中，該組合物(例如抗體、多核苷酸、載體、宿主細胞或醫藥組合物)經靜脈內、經腹膜內、經肌肉內、經動脈內、經鞘內、經淋巴內、經病灶內、經囊內、經眶內、經心臟內、經皮內、經氣管、經皮下、經表皮下、經關節內、經囊下、經蛛網膜下、經脊柱內、經硬膜外、經胸骨內、經局部、經表皮、經黏膜投與或其任何組合。在一些態樣中，該組合物經靜脈內投與。在一些態樣中，該組合物經皮下投與。The compositions of the present invention can be administered using any pharmaceutically acceptable route. In some aspects, the composition (e.g., antibody, polynucleotide, vector, host cell, or pharmaceutical composition) is administered intravenously, intraperitoneally, intramuscularly, intraarterially, intrathecally, intralymphatically, intralesionally, intracapsularly, intraorbitally, intracardially, intradermally, intratracheally, subcutaneously, subcutaneously, intraarticularly, subcapsularly, subarachnoidally, intraspinally, epidurally, intrasternally, topically, epidermally, transmucosally, or any combination thereof. In some aspects, the composition is administered intravenously. In some aspects, the composition is administered subcutaneously.

在某些態樣中，該方法減小該個體中之癌症之尺寸，例如腫瘤之尺寸。在一些態樣中，該癌症之尺寸減小至少約5%、至少約10%、至少約15%、至少約20%、至少約25%、至少約30%、至少約35%、至少約40%、至少約45%、至少約50%、至少約60%、至少約70%、至少約80%或至少約90%。In some aspects, the method reduces the size of a cancer, such as a tumor, in the individual. In some aspects, the size of the cancer is reduced by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%.

在一些態樣中，該方法增加該個體之總存活期。在一些態樣中，該總存活期相對於患有相同癌症但經不同療法治療之個體之平均總存活期增加。在某些態樣中，該總存活期增加至少約10%、至少約20%、至少約25%、至少約50%、至少約2倍、至少約3倍、至少約5倍。在一些態樣中，該總存活期增加至少約一個月、至少約2個月、至少約3個月、至少約4個月、至少約5個月、至少約6個月、至少約7個月、至少約8個月、至少約9個月、至少約10個月、至少約1個月、至少約12個月、至少約15個月、至少約18個月、至少約21個月、至少約2年、至少約3年、至少約4年、至少約5年或至少約10年。In some aspects, the method increases the overall survival of the individual. In some aspects, the overall survival is increased relative to the average overall survival of individuals with the same cancer but treated with different therapies. In certain aspects, the overall survival increases by at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 2 times, at least about 3 times, at least about 5 times. In some aspects, the overall survival increases by at least about one month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 1 month, at least about 12 months, at least about 15 months, at least about 18 months, at least about 21 months, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, or at least about 10 years.

在一些態樣中，該方法增加該個體之無疾病進展存活期。在一些態樣中，該總存活期相對於患有相同癌症但經不同療法治療之個體之平均無疾病進展存活期增加。在某些態樣中，該無疾病進展存活期增加至少約10%、至少約20%、至少約25%、至少約50%、至少約2倍、至少約3倍、至少約5倍。在一些態樣中，該總存活期增加至少約一個月、至少約2個月、至少約3個月、至少約4個月、至少約5個月、至少約6個月、至少約7個月、至少約8個月、至少約9個月、至少約10個月、至少約1個月、至少約12個月、至少約15個月、至少約18個月、至少約21個月、至少約2年、至少約3年、至少約4年、至少約5年或至少約10年。In some aspects, the method increases the disease-free survival of the individual. In some aspects, the overall survival is increased relative to the average disease-free survival of individuals with the same cancer but treated with different therapies. In certain aspects, the disease-free survival is increased by at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 2 times, at least about 3 times, at least about 5 times. In some aspects, the overall survival is increased by at least about one month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 1 month, at least about 12 months, at least about 15 months, at least about 18 months, at least about 21 months, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, or at least about 10 years.

在一些態樣中，該方法增加該個體之客觀反應率。在某些態樣中，該方法誘導該個體中之完全反應。在某些態樣中，該方法誘導該個體中之部分反應。In some aspects, the method increases the objective response rate of the subject. In some aspects, the method induces a complete response in the subject. In some aspects, the method induces a partial response in the subject.

在一些態樣中，該方法包括投與本文揭示之抗CD161抗體(或多核苷酸、載體或宿主細胞)及第二療法。在一些態樣中，該第二療法在該抗CD161抗體之前投與。在一些態樣中，該第二療法在該抗CD161抗體之後投與。在一些態樣中，該第二療法與該抗CD161抗體同時投與。在某些態樣中，該抗CD161抗體及該第二療法係分開投與。在其他態樣中，該抗CD161抗體及該第二療法以單一調配物投與。In some aspects, the method includes administering an anti-CD161 antibody (or polynucleotide, vector or host cell) disclosed herein and a second therapy. In some aspects, the second therapy is administered before the anti-CD161 antibody. In some aspects, the second therapy is administered after the anti-CD161 antibody. In some aspects, the second therapy is administered simultaneously with the anti-CD161 antibody. In some aspects, the anti-CD161 antibody and the second therapy are administered separately. In other aspects, the anti-CD161 antibody and the second therapy are administered with a single formulation.

該第二療法可係任一其他療法。在一些態樣中，該第二療法包含免疫療法。在一些態樣中，該第二療法包含化學療法。在一些態樣中，該第二療法包含放射療法。在一些態樣中，該第二療法包含外科手術。在一些態樣中，該第二療法包含投與第二治療劑。The second treatment can be any other treatment. In some aspects, the second treatment comprises immunotherapy. In some aspects, the second treatment comprises chemotherapy. In some aspects, the second treatment comprises radiation therapy. In some aspects, the second treatment comprises surgery. In some aspects, the second treatment comprises administration of a second therapeutic agent.

在一些態樣中，該一或多種另外治療劑係PD-1拮抗劑、TIM-3抑制劑、LAG-3抑制劑、TIGIT抑制劑、CD112R抑制劑、TAM抑制劑、STING促效劑、4-1BB促效劑或其組合。在一些態樣中，該一或多種另外治療劑係CD39拮抗劑、CD73拮抗劑、CCR8拮抗劑或其組合。在一些態樣中，該抗CD73係揭示於例如美國公開案第2019/0031766 A1號中之任一抗CD73抗體，該案之全文以引用之方式併入本文中。在一些態樣中，該抗CD39係揭示於例如國際公開案第WO 2019/178269 A2號中之任一抗CD39抗體，該案之全文以引用之方式併入本文中。In some embodiments, the one or more additional therapeutic agents are PD-1 antagonists, TIM-3 inhibitors, LAG-3 inhibitors, TIGIT inhibitors, CD112R inhibitors, TAM inhibitors, STING agonists, 4-1BB agonists, or combinations thereof. In some embodiments, the one or more additional therapeutic agents are CD39 antagonists, CD73 antagonists, CCR8 antagonists, or combinations thereof. In some embodiments, the anti-CD73 is any anti-CD73 antibody disclosed in, for example, U.S. Publication No. 2019/0031766 A1, the entirety of which is incorporated herein by reference. In some aspects, the anti-CD39 is any anti-CD39 antibody disclosed in, for example, International Publication No. WO 2019/178269 A2, the entire text of which is incorporated herein by reference.

在某些態樣中，該第二治療劑包含第二抗體。在一些態樣中，該第二治療劑包含有效量之特異性結合選自下列之蛋白質之抗體：誘導型T細胞共刺激因子(ICOS)、CD137 (4-1BB)、CD134 (OX40)、NKG2A、CD27、糖皮質激素誘導之TNFR相關蛋白(GITR)、皰疹病毒介入介質(HVEM)、程式化死亡-1 (PD-1)、程式化死亡配位體-1 (PD-L1)、CTLA-4、B及T淋巴細胞衰減蛋白(BTLA)、T細胞免疫球蛋白及黏蛋白域-3 (TIM-3)、淋巴細胞活化基因-3 (LAG-3)、腺苷A2a受體(A2aR)、殺手細胞凝集素樣受體G1 (KLRG-1)、天然殺手細胞受體2B4 (CD244)、CD160、具有Ig及ITIM域之T細胞免疫受體(TIGIT)、T細胞活化之V域Ig抑制因子之受體(VISTA)、NKG2a、KIR、TGFβ、IL-10、IL-8、B7-H4、Fas配位體、CXCR4、間皮素、CEACAM-1、CD96、CD52、HER2、及其任何組合。In some embodiments, the second therapeutic agent comprises a second antibody. In some embodiments, the second therapeutic agent comprises an effective amount of an antibody that specifically binds to a protein selected from the following: inducible T cell co-stimulator (ICOS), CD137 (4-1BB), CD134 (OX40), NKG2A, CD27, glucocorticoid-induced TNFR-related protein (GITR), herpes virus-mediated mediator (HVEM), programmed death-1 (PD-1), programmed death ligand-1 (PD-L1), CTLA-4, B and T lymphocyte attenuation protein (BTLA), T cell immunoglobulin and mucin domain-3 (TIM-3), lymphocyte activation gene-3 (LAG-3), adenosine A2a receptor (A2aR), killer cell lectin-like receptor G1 (KLRG-1), natural killer cell receptor 2B4 (CD244), CD160, T cell immune receptor with Ig and ITIM domains (TIGIT), receptor for V-domain Ig inhibitory factor of T cell activation (VISTA), NKG2a, KIR, TGFβ, IL-10, IL-8, B7-H4, Fas ligand, CXCR4, mesothelin, CEACAM-1, CD96, CD52, HER2, and any combination thereof.

本文描述之抗CD161抗體或其抗原結合部分可取代或增強先前或目前投與之療法。例如，在用抗CD161抗體或其抗原結合部分治療後，可停止或減少該一或多種另外治療劑之投與，例如，以較低濃度投與。在一些態樣中，可維持先前療法之投與。在一些態樣中，將維持先前療法直至抗CD161抗體之濃度達到足以提供治療效應之濃度。The anti-CD161 antibodies or antigen binding portions thereof described herein can replace or enhance a previously or currently administered therapy. For example, after treatment with an anti-CD161 antibody or antigen binding portion thereof, the administration of one or more additional therapeutic agents can be stopped or reduced, for example, administered at a lower concentration. In some aspects, the administration of the previous therapy can be maintained. In some aspects, the previous therapy will be maintained until the concentration of the anti-CD161 antibody reaches a concentration sufficient to provide a therapeutic effect.

本文描述之抗CD161抗體或其抗原結合部分可用作診斷劑。在一些態樣中，該抗CD161抗體用於識別適合於免疫療法之個體。在一些態樣中，該抗CD161抗體用於識別在NK細胞中具有CD161之表現的個體。The anti-CD161 antibodies or antigen-binding portions thereof described herein can be used as diagnostic agents. In some embodiments, the anti-CD161 antibodies are used to identify individuals suitable for immunotherapy. In some embodiments, the anti-CD161 antibodies are used to identify individuals with expression of CD161 in NK cells.

在一些態樣中，該一或多種另外治療劑係PD-1拮抗劑。在一些態樣中，該PD-1拮抗劑選自由下列組成之群：PDR001、納武單抗(nivolumab)、帕姆單抗(pembrolizumab)、彼地利株單抗(pidilizumab)、MEDI0680、REGN2810、TSR-042、PF-06801591及AMP-224。在某些態樣中，該一或多種另外治療劑係PD-L1抑制劑。在一些態樣中，該PD-L1抑制劑選自由下列組成之群：FAZ053、阿特珠單抗(Atezolizumab)、阿維單抗(Avelumab)、度伐魯單抗(Durvalumab)及BMS-936559。在一些態樣中，本發明提供一種增強抗PD-1抗體之一或多種活性(例如，增強PD-1介導之細胞激素分泌；增強抗PD-1介導之TNFα分泌；增強暴露至抗PD-1抗體之細胞之抗PD-1介導之IL-6分泌)之方法，該方法包括將細胞與抗PD-1抗體同時或依序暴露至本發明提供之抗體或其抗原結合部分，由此增強該抗PD1抗體之一或多種活性。In some embodiments, the one or more additional therapeutic agents are PD-1 antagonists. In some embodiments, the PD-1 antagonists are selected from the group consisting of PDR001, nivolumab, pembrolizumab, pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, and AMP-224. In certain embodiments, the one or more additional therapeutic agents are PD-L1 inhibitors. In some embodiments, the PD-L1 inhibitors are selected from the group consisting of FAZ053, Atezolizumab, Avelumab, Durvalumab, and BMS-936559. In some aspects, the present invention provides a method for enhancing one or more activities of an anti-PD-1 antibody (e.g., enhancing PD-1-mediated cytokine secretion; enhancing anti-PD-1-mediated TNFα secretion; enhancing anti-PD-1-mediated IL-6 secretion of cells exposed to anti-PD-1 antibodies), the method comprising exposing cells and anti-PD-1 antibodies to an antibody or an antigen-binding portion thereof provided by the present invention simultaneously or sequentially, thereby enhancing one or more activities of the anti-PD1 antibody.

在一些態樣中，該一或多種另外治療劑係舒尼替尼(Sunitinib) (Sutent^®)、卡博替尼(Cabozantinib) (CABOMETYX^®)、阿西替尼(Axitinib) (INLYTA^®)、樂伐替尼(Lenvatinib) (LENVIMA^®)、依維莫司(Everolimus) (AFINITOR^®)、貝伐單抗(Bevacizumab) (AVASTIN^®)、艾帕斯塔(epacadostat)、NKTR-214 (CD-122偏向促效劑(CD-122-biased agonist))、替沃贊尼(tivozanib) (FOTIVDA^®)、阿貝新司他(abexinostat)、易普利單抗(Ipilimumab) (YERVOY^®)、曲美目單抗(tremelimumab)、帕唑帕尼(Pazopanib) (VOTRIENT^®)、索拉非尼(Sorafenib) (NEXAVAR^®)、替西羅莫司(Temsirolimus) (TORISEL^®)、雷莫蘆單抗(Ramucirumab) (CYRAMZA^®)、尼拉帕尼(niraparib)、沃利替尼(savolitinib)、伏羅尼布(vorolanib) (X-82)、瑞格菲尼(Regorafenib) (STIVARGO^®)、道那菲尼(Donafenib) (多激酶抑制劑)、卡瑞利珠單抗(Camrelizumab) (SHR-1210)、派替莫金德瓦維克(pexastimogene devacirepvec) (JX-594)、雷莫蘆單抗(Ramucirumab) (CYRAMZA^®)、阿帕替尼(apatinib) (YN968D1)、囊封的多柔比星(encapsulated doxorubicin) (THERMODOX^®)、提瓦替尼(Tivantinib) (ARQ197)、ADI-PEG 20、畢尼替尼(binimetinib)、甲磺酸阿帕替尼(apatinib mesylate)、尼达尼布(nintedanib)、利麗單抗(lirilumab)、納武單抗(OPDIVO^®)、帕姆單抗(Pembrolizumab) (KEYTRUDA^®)、阿特珠單抗(TECENTRIQ^®)、阿維單抗(BAVENCIO^®)、度伐魯單抗(IMFIMZI^®)、西米普利單抗-rwlc (Cemiplimab-rwlc) (LIBTAYO^®)、替雷利珠單抗(tislelizumab)及/或斯巴達珠單抗(spartalizumab)。In some embodiments, the one or more additional therapeutic agents are sunitinib (Sutent®), cabozantinib (^CABOMETYX® ), axitinib (^INLYTA® ), lenvatinib (LENVIMA®), everolimus (AFINITOR®), bevacizumab (AVASTIN®), epacadostat, NKTR-214 (CD-122-biased agonist), tivozanib (^FOTIVDA® ), abexinostat, ipilimumab (^YERVOY® ), tadalafil (Tadalafil®), selegiline (Sutent®), tadalafil (Tadalafil®), selegiline (Sutent®⁾ , selegiline (Sutent®), selegiline (Sutent®), selegiline (Sutent®), selegiline (^Sutent® ), selegiline (Sutent®), selegiline (Sutent®), selegiline (Sutent®), selegiline (Sutent®), selegiline (Sutent®), selegiline (Sutent®), selegiline (Sutent®), selegiline (Sutent®), selegiline (Sutent®), selegiline (^Sutent® ), selegiline (Sule®), selegiline (Sule®), selegiline (Sule®), selegiline (Sule®), selegiline (Sule®), selegiline (Sule®), selegiline (^Sule ), tremelimumab, pazopanib (VOTRIENT^{® ), sorafenib (NEXAVAR ®} ), temsirolimus (TORISEL ® ), ramucirumab (CYRAMZA^®) , niraparib, savolitinib, vorolanib (X-82), regorafenib (STIVARGO^® ), donafenib (multikinase inhibitor), camrelizumab (SHR-1210), pexastimogene devacirepvec (JX-594), ramucirumab (^CYRAMZA® ), apatinib (YN968D1), encapsulated doxorubicin (THERMODOX^® ), tivinib (ARQ197), ADI-PEG 20, binimetinib, apatinib mesylate, nintedanib, lirilumab, OPDIVO^® , pembrolizumab (KEYTRUDA^® ), TECENTRIQ^® , BAVENCIO^® , IMFIMZI^® , Cemiplimab-rwlc (LIBTAYO^® ), tislelizumab and/or spartalizumab.

在一些態樣中，該一或多種另外治療劑係TIM-3抑制劑，視情況，其中該TIM-3抑制劑係MGB453或TSR-022。In some aspects, the one or more additional therapeutic agents is a TIM-3 inhibitor, optionally wherein the TIM-3 inhibitor is MGB453 or TSR-022.

在一些態樣中，該一或多種另外治療劑係LAG-3抑制劑，視情況，其中該LAG-3抑制劑選自由LAG525、BMS-986016及TSR-033組成之群。In some aspects, the one or more additional therapeutic agents is a LAG-3 inhibitor, optionally, wherein the LAG-3 inhibitor is selected from the group consisting of LAG525, BMS-986016, and TSR-033.

在一些態樣中，該一或多種另外治療劑係TIGIT抑制劑。在一些態樣中，該一或多種另外治療劑係CD112R抑制劑。在一些態樣中，該一或多種另外治療劑係TAM (Axl、Mer、Tyro)抑制劑。在一些態樣中，該一或多種另外治療劑係STING促效劑。在一些態樣中，該一或多種另外治療劑係4-1BB促效劑。In some embodiments, the one or more additional therapeutic agents are TIGIT inhibitors. In some embodiments, the one or more additional therapeutic agents are CD112R inhibitors. In some embodiments, the one or more additional therapeutic agents are TAM (Axl, Mer, Tyro) inhibitors. In some embodiments, the one or more additional therapeutic agents are STING agonists. In some embodiments, the one or more additional therapeutic agents are 4-1BB agonists.

在一些態樣中，該一或多種另外治療劑係酪胺酸激酶抑制劑、靶向腺苷軸線之藥劑(例如CD39拮抗劑、CD73拮抗劑或A2AR、A2BR或雙重A2AR/A2BR拮抗劑)、CCR8拮抗劑、CTLA4拮抗劑、VEG-F抑制劑或其組合。 III.A.1. 與化療劑之組合In some embodiments, the one or more additional therapeutic agents are tyrosine kinase inhibitors, agents targeting the adenosine axis (e.g., CD39 antagonists, CD73 antagonists, or A2AR, A2BR, or dual A2AR/A2BR antagonists), CCR8 antagonists, CTLA4 antagonists, VEG-F inhibitors, or combinations thereof.III.A.1. Combinations with chemotherapeutic agents

適合於與本發明組合物組合及/或共同投與之化療劑包括(例如)：紫杉醇(taxol)、細胞鬆弛素B、短桿菌肽D、溴化乙錠、吐根素(emetine)、絲裂黴素、依託泊苷(etoposide)、替尼泊苷(tenoposide)、長春新鹼(vincristine)、長春花鹼(vinblastine)、秋水仙鹼(colchicin)、多柔比星、道諾黴素(daunorubicin)、二羥基蒽二酮(dihydroxyanthrancindione)、米托蒽醌(mitoxantrone)、米沙黴素(mithramycin)、放線菌素D (actinomycin D)、1-去氫睾固酮、糖皮質激素、普魯卡因、地卡因(tetracaine)、利多卡因(lidocaine)、普萘洛爾(propranolol)及嘌呤黴素及其類似者或同源物。其他藥劑包括(例如)抗代謝物(例如胺甲喋呤(methotrexate)、6-巰基嘌呤、6-硫鳥嘌呤、阿糖胞苷(cytarabine)、5-氟尿嘧啶達卡巴嗪(decarbazine))、烷基化劑(例如氮芥(mechlorethamine)、硫代TEPA、苯丁酸氮芥(chlorambucil)、美法侖(melphalan)、卡莫司汀(carmustine) (BSNU)、洛莫司汀(lomustine) (CCNU)、環磷醯胺、白消安(busulfan)、二溴甘露醇、鏈佐黴素(streptozotocin)、絲裂黴素C、順式-二氯二胺鉑(II)(DDP)、丙卡巴肼(procarbazine)、六甲蜜胺(altretamine)、順鉑、卡鉑(carboplatin)、奧沙利鉑(oxaliplatin)、奈達鉑(nedaplatin)、賽特鉑(satraplatin)或四硝酸特瑞鉑(triplatin tetranitrate))、蒽環黴素(例如道諾黴素(先前為道諾黴素(daunomycin))及多柔比星)、抗生素(例如更生黴素(dactinomcin) (先前為放線菌素)、博來黴素(bleomycin)、米沙黴素、及安曲黴素(anthramycin) (AMC))、及抗有絲分裂劑(例如長春新鹼及長春花鹼)及替莫唑胺(temozolomide)。 III.A.2. 與PD-1/PD-L1拮抗劑之組合Chemotherapeutic agents suitable for combination and/or co-administration with the composition of the present invention include, for example, paclitaxel, cytochalasin B, bacitracin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxyanthrancindione, mitoxantrone, mithramycin, actinomycin D, d-hydroxy- ... D), 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol and puromycin and their analogs or homologs. Other agents include, for example, anti-metabolites (e.g., methotrexate, 6-hydroxypurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thio-TEPA, chlorambucil, melphalan, carmustine (BSNU), lomustine) (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, cis-dichlorodiamine platinum (II) (DDP), procarbazine, altretamine, cis-platinum, carboplatin, oxaliplatin, nedaplatin, satraplatin, or triplatin tetranitrate), anthracyclines (e.g., daunomycin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomcin) (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC), and antimitotic agents (such as vincristine and vinblastine) and temozolomide.III.A.2. Combinations with PD-1/PD-L1 antagonists

在一些態樣中，將本發明提供之抗CD161抗體或其抗原結合部分與一或多種PD-1拮抗劑組合(例如組合投與)，該一或多種PD-1拮抗劑特異性結合至人類PD-1或PD-L1且抑制PD-1/PD-L1生物活性及/或藉由人類PD-1/PD-L1信號傳導介導之下游路徑及/或細胞製程或其他人類PD-1/PD-L1介導之功能。In some aspects, the anti-CD161 antibodies or antigen-binding portions thereof provided herein are combined (e.g., administered in combination) with one or more PD-1 antagonists that specifically bind to human PD-1 or PD-L1 and inhibit PD-1/PD-L1 biological activity and/or downstream pathways mediated by human PD-1/PD-L1 signaling and/or cellular processes or other human PD-1/PD-L1-mediated functions.

因此，本文提供PD-1拮抗劑，其直接或變構阻斷、拮抗、遏制(suppress)、抑制(inhibit)或減少PD-1/PD-L1生物活性，包括藉由PD-1/PD-L1信號傳導介導之下游路徑及/或細胞過程，諸如受體結合及/或引發對PD-1/PD-L1之細胞反應。本文亦提供PD-1拮抗劑，其減少由細胞或個體產生之人類PD-1或PD-L1之數量或量。Therefore, provided herein are PD-1 antagonists that directly or allosterically block, antagonize, suppress, inhibit or reduce PD-1/PD-L1 biological activity, including downstream pathways and/or cellular processes mediated by PD-1/PD-L1 signaling, such as receptor binding and/or induced cellular responses to PD-1/PD-L1. Also provided herein are PD-1 antagonists that reduce the number or amount of human PD-1 or PD-L1 produced by cells or individuals.

在一些態樣中，本發明提供結合人類PD-1且阻止、抑制或減少PD-L1結合至PD-1之PD-1拮抗劑。在一些態樣中，該PD-1拮抗劑結合至編碼PD-1或PD-L1之mRNA且阻止轉譯。在一些態樣中，該PD-1拮抗劑結合至編碼PD-1或PD-L1之mRNA且導致降解及/或轉換(turnover)。In some aspects, the present invention provides a PD-1 antagonist that binds to human PD-1 and prevents, inhibits or reduces the binding of PD-L1 to PD-1. In some aspects, the PD-1 antagonist binds to mRNA encoding PD-1 or PD-L1 and prevents translation. In some aspects, the PD-1 antagonist binds to mRNA encoding PD-1 or PD-L1 and causes degradation and/or turnover.

在一些態樣中，該PD-1拮抗劑抑制PD-1信號傳導或功能。在一些態樣中，該PD-1拮抗劑阻斷PD-1與PD-L1、PD-L2或與PD-L1及PD-L2二者之結合。在一些態樣中，該PD-1拮抗劑阻斷PD-1與PD-L1之結合。在一些態樣中，該PD-1拮抗劑阻斷PD-1與PD-L2之結合。在一些態樣中，該PD-1拮抗劑阻斷PD-1與PD-L1及PD-L2之結合。在一些態樣中，該PD-1拮抗劑特異性結合PD-1。在一些態樣中，該PD-1拮抗劑特異性結合PD-L1。在一些態樣中，該PD-1拮抗劑特異性結合PD-L2。In some embodiments, the PD-1 antagonist inhibits PD-1 signaling or function. In some embodiments, the PD-1 antagonist blocks the binding of PD-1 to PD-L1, PD-L2, or both PD-L1 and PD-L2. In some embodiments, the PD-1 antagonist blocks the binding of PD-1 to PD-L1. In some embodiments, the PD-1 antagonist blocks the binding of PD-1 to PD-L2. In some embodiments, the PD-1 antagonist blocks the binding of PD-1 to PD-L1 and PD-L2. In some embodiments, the PD-1 antagonist specifically binds to PD-1. In some embodiments, the PD-1 antagonist specifically binds to PD-L1. In some aspects, the PD-1 antagonist specifically binds PD-L2.

在一些態樣中，PD-1拮抗劑抑制PD-1與其同源配位體之結合。在一些態樣中，該PD-1拮抗劑抑制PD-1與PD-L1、PD-1與PD-L2、或PD-1與PD-L1及PD-L2二者之結合。在一些態樣中，該PD-1拮抗劑不抑制PD-1與其同源配位體之結合。In some aspects, the PD-1 antagonist inhibits the binding of PD-1 to its cognate ligand. In some aspects, the PD-1 antagonist inhibits the binding of PD-1 to PD-L1, PD-1 to PD-L2, or PD-1 to both PD-L1 and PD-L2. In some aspects, the PD-1 antagonist does not inhibit the binding of PD-1 to its cognate ligand.

在一些態樣中，該PD-1拮抗劑係分離的抗體(mAb)或其抗原結合片段，其特異性結合至PD-1或PD-L1。在一些態樣中，該PD-1拮抗劑係抗體或其抗原結合片段，其特異性結合至人類PD-1。在一些態樣中，該PD-1拮抗劑係抗體或其抗原結合片段，其特異性結合至人類PD-L1。在一些態樣中，該PD-1拮抗劑係抗體或其抗原結合片段，其結合至人類PD-L1且抑制PD-L1與PD-1之結合。在一些態樣中，該PD-1拮抗劑係抗體或其抗原結合片段，其結合至人類PD-1且抑制PD-L1與PD-1之結合。In some aspects, the PD-1 antagonist is an isolated antibody (mAb) or an antigen-binding fragment thereof that specifically binds to PD-1 or PD-L1. In some aspects, the PD-1 antagonist is an antibody or an antigen-binding fragment thereof that specifically binds to human PD-1. In some aspects, the PD-1 antagonist is an antibody or an antigen-binding fragment thereof that specifically binds to human PD-L1. In some aspects, the PD-1 antagonist is an antibody or an antigen-binding fragment thereof that specifically binds to human PD-L1. In some aspects, the PD-1 antagonist is an antibody or an antigen-binding fragment thereof that binds to human PD-L1 and inhibits the binding of PD-L1 to PD-1. In some aspects, the PD-1 antagonist is an antibody or an antigen-binding fragment thereof that binds to human PD-1 and inhibits the binding of PD-L1 to PD-1.

抑制或破壞PD-1與其配位體PD-L1及PD-L2中之一者或二者之間的相互作用之幾種免疫檢查點拮抗劑正處於臨床開發中或目前臨床醫生可用於治療癌症。Several immune checkpoint antagonists that inhibit or disrupt the interaction between PD-1 and one or both of its ligands PD-L1 and PD-L2 are in clinical development or currently available to clinicians for the treatment of cancer.

在本發明提供之組合物、方法及用途中任一者中可包含PD-1拮抗劑之抗人類PD-1抗體或其抗原結合片段之實例包括(但不限於)：KEYTRUDA^®(帕姆單抗、MK-3475、h409A11；參見US8952136、US8354509、US8900587及EP2170959，其等均以全文引用之方式包括在本文中；Merck)、OPDIVO^®(納武單抗、BMS-936558、MDX-1106、ONO-4538；參見US7595048、US8728474、US9073994、US9067999、EP1537878、US8008449、US8779105及EP2161336，其等均以全文引用之方式包括在本文中；Bristol Myers Squibb)、MEDI0680 (AMP-514)、BGB-A317及BGB-108 (BeiGene)、244C8及388D4 (參見WO2016106159，該案以全文引用之方式併入本文中；Enumeral Biomedical)、PDR001 (Novartis)及REGN2810 (Regeneron)。因此，在一些態樣中，該PD-1拮抗劑係帕姆單抗。在一些態樣中，該PD-1拮抗劑係納武單抗。Examples of anti-human PD-1 antibodies or antigen-binding fragments thereof that may comprise PD-1 antagonists in any of the compositions, methods and uses provided herein include, but are not limited to:^KEYTRUDA® (pembrolizumab, MK-3475, h409A11; see US8952136, US8354509, US8900587 and EP2170959, all of which are incorporated herein by reference in their entirety; Merck),^OPDIVO® (nivolumab, BMS-936558, MDX-1106, ONO-4538; see US7595048, US8728474, US9073994, US9067999, EP1537878, US8008449, US8779105 and EP2161336, all of which are incorporated herein by reference in their entirety; Bristol Myers Squibb), MEDI0680 (AMP-514), BGB-A317 and BGB-108 (BeiGene), 244C8 and 388D4 (see WO2016106159, which is incorporated herein by reference in its entirety; Enumeral Biomedical), PDR001 (Novartis) and REGN2810 (Regeneron). Thus, in some aspects, the PD-1 antagonist is pembrolizumab. In some aspects, the PD-1 antagonist is nivolumab.

在本發明提供之組合物、方法及用途中任一者中可包含PD-L1拮抗劑之抗人類PD-1抗體或其抗原結合片段之實例包括(但不限於)：BAVENCIO^®(阿維單抗、MSB0010718C，參見WO2013/79174，該案以全文引用之方式併入本文中；Merck/Pfizer)、IMFINZI^®(度伐魯單抗、MEDI4736)、TECENTRIQ^®(阿特珠單抗、MPDL3280A、RG7446；參見WO2010/077634，該案以全文引用之方式併入本文中；Roche)、MDX-1105 (BMS-936559、12A4；參見US7943743及WO2013/173223，其等皆以全文引用之方式併入本文中；Medarex/BMS)、及FAZ053 (Novartis)。因此，在一些態樣中，該PD-1拮抗劑係阿維單抗。在一些態樣中，該PD-1拮抗劑係度伐魯單抗。在一些態樣中，該PD-1拮抗劑係阿特珠單抗。Examples of anti-human PD-1 antibodies or antigen-binding fragments thereof that may comprise a PD-L1 antagonist in any of the compositions, methods, and uses provided herein include, but are not limited to:^BAVENCIO® (avelumab, MSB0010718C, see WO2013/79174, which is incorporated herein by reference in its entirety; Merck/Pfizer), IMFINZI® (durvalumab, MEDI4736),^TECENTRIQ® (atezolizumab, MPDL3280A, RG7446; see WO2010/077634, which is incorporated herein by reference in its entirety; Roche), MDX^- 1105® (durvalumab, MEDI4736), and TECENTRIQ® (atezolizumab, MPDL3280A, RG7446; see WO2010/077634, which is incorporated herein by reference in its entirety; Roche). (BMS-936559, 12A4; see US7943743 and WO2013/173223, all of which are incorporated herein by reference in their entirety; Medarex/BMS), and FAZ053 (Novartis). Thus, in some aspects, the PD-1 antagonist is avelumab. In some aspects, the PD-1 antagonist is durvalumab. In some aspects, the PD-1 antagonist is atezolizumab.

在一些態樣中，該PD-1拮抗劑係特異性結合至人類PD-1或人類PD-L1之免疫黏附素，例如含有融合至恆定區(諸如免疫球蛋白分子之Fc區)之PD-L1或PD-L2之細胞外或PD-1結合部分之融合蛋白。特異性結合至PD-1之免疫黏附分子之實例描述於WO2010/027827及WO2011/066342中，該兩案以全文引用之方式併入本文中。在一些態樣中，該PD-1拮抗劑係AMP-224 (亦稱為B7-DCIg)，其係特異性結合至人類PD-1之PD-L2-FC融合蛋白。In some aspects, the PD-1 antagonist is an immunoadhesin that specifically binds to human PD-1 or human PD-L1, such as a fusion protein containing an extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a homeostatic region (such as the Fc region of an immunoglobulin molecule). Examples of immunoadhesion molecules that specifically bind to PD-1 are described in WO2010/027827 and WO2011/066342, both of which are incorporated herein by reference in their entirety. In some aspects, the PD-1 antagonist is AMP-224 (also known as B7-DCIg), which is a PD-L2-FC fusion protein that specifically binds to human PD-1.

一般技術者應理解，結合至PD-1或PD-L1且破壞PD-1/PD-L1信號傳導路徑之任一PD-1拮抗劑適合於本文揭示之組合物、方法及用途。It will be understood by those of ordinary skill in the art that any PD-1 antagonist that binds to PD-1 or PD-L1 and disrupts the PD-1/PD-L1 signaling pathway is suitable for the compositions, methods, and uses disclosed herein.

在一些態樣中，PD-1/PD-L1拮抗劑係小分子、核酸、肽、肽模擬物、蛋白質、碳水化合物、碳水化合物衍生物或醣聚合物(glycopolymer)。示例性小分子PD-1抑制劑描述於Zhan等人(2016) Drug Discov Today 21(6):1027-1036中。 III.A.3. 與TIM-3抑制劑之組合In some embodiments, the PD-1/PD-L1 antagonist is a small molecule, a nucleic acid, a peptide, a peptide mimetic, a protein, a carbohydrate, a carbohydrate derivative, or a glycopolymer. Exemplary small molecule PD-1 inhibitors are described in Zhan et al. (2016) Drug Discov Today 21(6):1027-1036.III.A.3. Combination with TIM-3 inhibitors

在一些態樣中，將本發明提供之抗CD161抗體或其抗原結合部分與TIM-3抑制劑組合(例如組合投與)。該TIM-3抑制劑可係抗體、其抗原結合片段、免疫黏附素、融合蛋白或寡肽。在一些態樣中，該TIM-3抑制劑選自MGB453 (Novartis)、TSR-022 (Tesaro)或LY3321367 (Eli Lilly)。在一些態樣中，將該抗CD161抗體或其抗原結合部分與MGB453組合投與。在一些態樣中，將該抗CD161抗體或其抗原結合部分與TSR-022組合投與。 III.A.4. 與LAG-3抑制劑之組合In some embodiments, the anti-CD161 antibody or its antigen binding portion provided by the present invention is combined with a TIM-3 inhibitor (e.g., administered in combination). The TIM-3 inhibitor can be an antibody, an antigen binding fragment thereof, an immunoadhesin, a fusion protein, or an oligopeptide. In some embodiments, the TIM-3 inhibitor is selected from MGB453 (Novartis), TSR-022 (Tesaro), or LY3321367 (Eli Lilly). In some embodiments, the anti-CD161 antibody or its antigen binding portion is administered in combination with MGB453. In some embodiments, the anti-CD161 antibody or its antigen binding portion is administered in combination with TSR-022.III.A.4. Combination with LAG-3 inhibitors

在一些態樣中，將本發明提供之抗CD161抗體或其抗原結合部分與LAG-3抑制劑組合(例如組合投與)。該LAG-3抑制劑可係抗體、其抗原結合片段、免疫黏附素、融合蛋白或寡肽。在一些態樣中，該LAG-3抑制劑選自LAG525 (Novartis)、BMS-986016 (Bristol-Myers Squibb)、TSR-033 (Tesaro)、MK-4280 (Merck及Co)或REGN3767 (Regeneron)。 III.A.5. 其他組合In some embodiments, the anti-CD161 antibody or antigen-binding portion thereof provided by the present invention is combined with a LAG-3 inhibitor (e.g., administered in combination). The LAG-3 inhibitor can be an antibody, an antigen-binding fragment thereof, an immunoadhesin, a fusion protein, or an oligopeptide. In some embodiments, the LAG-3 inhibitor is selected from LAG525 (Novartis), BMS-986016 (Bristol-Myers Squibb), TSR-033 (Tesaro), MK-4280 (Merck and Co), or REGN3767 (Regeneron).III.A.5. Other combinations

在一些態樣中，將本發明提供之抗CD161抗體或其抗原結合部分與TIGIT抑制劑、激酶抑制劑(例如酪胺酸激酶抑制劑(TKI))、CD112R抑制劑、TAM受體抑制劑、STING促效劑及/或4-1BB促效劑或其組合組合(例如組合投與)。在一些態樣中，將本發明提供之抗CD161抗體或其抗原結合部分與酪胺酸激酶抑制劑、靶向腺苷軸線之藥劑(例如CD39拮抗劑、CD73拮抗劑或A2AR、A2BR或雙重A2AR/A2BR拮抗劑)、CCR8拮抗劑、CTLA4拮抗劑、VEG-F抑制劑或其組合組合(例如組合投與)。 III.B. 工程化抗體之方法In some embodiments, the anti-CD161 antibody or its antigen-binding portion provided by the present invention is combined with a TIGIT inhibitor, a kinase inhibitor (e.g., a tyrosine kinase inhibitor (TKI)), a CD112R inhibitor, a TAM receptor inhibitor, a STING agonist and/or a 4-1BB agonist, or a combination thereof (e.g., administered in combination). In some embodiments, the anti-CD161 antibody or its antigen-binding portion provided by the present invention is combined with a tyrosine kinase inhibitor, an agent targeting the adenosine axis (e.g., a CD39 antagonist, a CD73 antagonist, or an A2AR, A2BR, or a dual A2AR/A2BR antagonist), a CCR8 antagonist, a CTLA4 antagonist, a VEG-F inhibitor, or a combination thereof (e.g., administered in combination).III.B. Methods of engineering antibodies

可使用本文揭示之具有VH及VL序列之抗CD161抗體來藉由修飾VH及/或VL序列或連接至其之恆定區來創建新的抗CD161抗體。因此，在本文描述之另一個態樣中，本文描述之抗CD161抗體之結構特徵用於創建結構相關之抗CD161抗體，其保留本文描述之抗CD161抗體之至少一種功能性質，諸如抑制人類CD161與CLEC2D之間之相互作用。例如，可將本文揭示之抗體之一或多個CDR區與已知框架區及/或其他CDR重組組合以創建本文描述之另外經重組工程化抗CD161抗體，如上文所討論。用於該工程化方法之起始材料係本文提供之VH及/或VL序列中之一者或多者、或其一或多個CDR區。為了創建該工程化抗體，不必實際製備(亦即以蛋白質表現)具有本文提供之VH及/或VL序列中之一者或多者、或其一或多個CDR區之抗體。相反，包含在該(等)序列中之資訊用作起始材料以創建衍生自該(等)原始序列之「第二代」序列且然後製備該(等)「第二代」序列且以蛋白質表現。Anti-CD161 antibodies with VH and VL sequences disclosed herein can be used to create new anti-CD161 antibodies by modifying VH and/or VL sequences or being connected to constant regions thereof. Therefore, in another aspect described herein, the structural features of the anti-CD161 antibodies described herein are used to create structurally related anti-CD161 antibodies that retain at least one functional property of the anti-CD161 antibodies described herein, such as inhibiting the interaction between human CD161 and CLEC2D. For example, one or more CDR regions of the antibodies disclosed herein can be recombined with known framework regions and/or other CDRs to create additional recombinant engineered anti-CD161 antibodies described herein, as discussed above. The starting material for the engineering method is one or more of the VH and/or VL sequences provided herein, or one or more CDR regions thereof. In order to create the engineered antibody, it is not necessary to actually prepare (i.e., express as a protein) an antibody having one or more of the VH and/or VL sequences provided herein, or one or more CDR regions thereof. Instead, the information contained in the sequence(s) is used as starting material to create a "second generation" sequence derived from the original sequence(s) and then the "second generation" sequence(s) is prepared and expressed as a protein.

此外，本文揭示之抗體可透過已知技術諸如親和力成熟來改良。親和力成熟係一種允許選擇以比起始抗體更大的親和力結合至抗原之衍生物抗體之技術。在一些態樣中，本文揭示之抗體用作起始抗體以進行親和力成熟。In addition, the antibodies disclosed herein can be improved by known techniques such as affinity maturation. Affinity maturation is a technique that allows the selection of derivative antibodies that bind to an antigen with greater affinity than the starting antibody. In some aspects, the antibodies disclosed herein are used as starting antibodies for affinity maturation.

因此，本文提供用於製備本文描述之抗CD161抗體之方法。 III.C. 抗體產生Thus, provided herein are methods for preparing the anti-CD161 antibodies described herein.III.C. Antibody Production

本文描述之抗CD161抗體可使用多種已知技術產生，諸如Kohler及Milstein,Nature256: 495 (1975)描述之標準體細胞雜交技術。雖然體細胞雜交程序常見，但原則上亦可採用用於產生單株抗體之其他技術，例如B淋巴細胞之病毒或致癌轉化、使用人類抗體基因庫之噬菌體展示技術。The anti-CD161 antibodies described herein can be produced using a variety of known techniques, such as the standard somatic cell hybridization technique described by Kohler and Milstein,Nature 256: 495 (1975). Although the somatic cell hybridization procedure is common, other techniques for producing monoclonal antibodies can also be used in principle, such as viral or oncogenic transformation of B lymphocytes, phage display technology using human antibody gene libraries.

在一些態樣中，用於製備雜交瘤之動物系統係鼠類系統。小鼠中之雜交瘤產生係一種非常完善的程序。用於分離用於融合之免疫化脾細胞之免疫方案及技術係此項技術中已知的。融合搭配物(例如鼠類骨髓瘤細胞)及融合程序亦係已知的。In some aspects, the animal system used to prepare the hybridoma is a murine system. Hybridoma generation in mice is a well-established procedure. Immunization protocols and techniques for isolating immunized spleen cells for fusion are known in the art. Fusion partners (e.g., murine myeloma cells) and fusion procedures are also known.

嵌合或人類化抗CD161抗體可基於如上所述製備的鼠類單株抗體之序列來製備。編碼重鏈及輕鏈免疫球蛋白之DNA可自所關注鼠類雜交瘤獲得且使用標準分子生物學技術工程化以含有非鼠類(例如人類)免疫球蛋白序列。例如，為了創建嵌合抗體，可使用此項技術中已知的方法將鼠類可變區連接至人類恆定區(參見，例如，Cabilly等人之美國專利第4,816,567號)。為了創建人類化抗體，可使用此項技術中已知的方法，將CDR區插入至人類框架中(參見，例如，Winter之美國專利第5,225,539號及Queen等人之美國專利第5,530,101號；第5,585,089號；第5,693,762號及第6,180,370號)。Chimeric or humanized anti-CD161 antibodies can be prepared based on the sequence of a mouse monoclonal antibody prepared as described above. DNA encoding heavy and light chain immunoglobulins can be obtained from a mouse hybridoma of interest and engineered to contain non-mouse (e.g., human) immunoglobulin sequences using standard molecular biology techniques. For example, to create chimeric antibodies, mouse variable regions can be linked to human constant regions using methods known in the art (see, e.g., U.S. Patent No. 4,816,567 to Cabilly et al.). To create humanized antibodies, the CDR regions can be inserted into a human framework using methods known in the art (see, e.g., U.S. Pat. Nos. 5,225,539 to Winter and 5,530,101; 5,585,089; 5,693,762; and 6,180,370 to Queen et al.).

在一些態樣中，本文描述之抗CD161抗體係人類單株抗體。此類針對CD161之人類單株抗體可使用攜載人類免疫系統而非小鼠系統之部分之轉殖基因或轉染色體小鼠來產生。此等轉殖基因或轉染色體小鼠包括本文中分別稱為HuMAb小鼠及KM小鼠之小鼠且在本文中統稱為「人類Ig小鼠」。In some aspects, the anti-CD161 antibodies described herein are human monoclonal antibodies. Such human monoclonal antibodies against CD161 can be produced using transgenic or transchromosomal mice carrying parts of the human immune system rather than the mouse system. These transgenic or transchromosomal mice include mice referred to herein as HuMAb mice and KM mice, respectively, and are collectively referred to herein as "human Ig mice".

HuMAb-mouse^®(Medarex, Inc.)含有編碼未重排的人類重鏈(μ及γ)及κ輕鏈免疫球蛋白序列之人類免疫球蛋白基因微基因座、以及將內源μ及κ鏈基因座去活化之靶向突變(參見，例如，Lonberg等人(1994)Nature368(6474): 856-859)。因此，小鼠展現小鼠IgM或κ之減少之表現，且回應於免疫，引入之人類重鏈及輕鏈轉殖基因經歷類別切換及體細胞突變以產生高親和力人類IgGK單株(Lonberg, N.等人(1994)，同上；綜述於Lonberg, N. (1994) Handbook of Experimental Pharmacology 113:49-101；Lonberg, N.及Huszar, D. (1995)Intern.Rev. Immunol. 13: 65-93、及Harding, F.及Lonberg, N. (1995)Ann. N.Y. Acad. Sci.764:536-546)中。HuMAb-^mouse® (Medarex, Inc.) contains a human immunoglobulin gene miniloci encoding unrearranged human heavy chain (μ and γ) and kappa light chain immunoglobulin sequences, and targeted mutations that inactivate the endogenous μ and kappa chain loci (see, e.g., Lonberg et al. (1994)Nature 368(6474): 856-859). Thus, the mice display reduced expression of mouse IgM or κ, and in response to immunization, the introduced human heavy and light chain transgenes undergo class switching and somatic mutagenesis to generate high affinity human IgGκ monoclonals (Lonberg, N. et al. (1994), supra; reviewed in Lonberg, N. (1994) Handbook of Experimental Pharmacology 113:49-101; Lonberg, N. and Huszar, D. (1995)Intern. Rev. Immunol . 13:65-93, and Harding, F. and Lonberg, N. (1995)Ann. NY Acad. Sci. 764:536-546).

在一些態樣中，使用在轉殖基因及轉染染色體上攜載人類免疫球蛋白序列之小鼠(諸如攜載人類重鏈轉殖基因及人類輕鏈轉染色體之小鼠)來產生本文描述之抗CD161抗體。此類小鼠(在本文中稱為「KM小鼠」)詳細描述於Ishida等人之PCT公開案WO 02/43478中。In some aspects, mice carrying human immunoglobulin sequences on transgenes and transfected chromosomes (such as mice carrying human heavy chain transgenes and human light chain transchromosomes) are used to generate anti-CD161 antibodies described herein. Such mice (referred to herein as "KM mice") are described in detail in PCT Publication WO 02/43478 by Ishida et al.

又此外，表現人類免疫球蛋白基因之替代轉殖基因動物系統係可用，其可用於刺激生成本文描述之抗CD161抗體。例如，可使用稱為Xenomouse (Abgenix, Inc.)之替代轉殖基因系統；此類小鼠描述於例如美國專利第5,939,598號中。In addition, alternative transgenic animal systems expressing human immunoglobulin genes are available, which can be used to stimulate the production of anti-CD161 antibodies described herein. For example, an alternative transgenic system called Xenomouse (Abgenix, Inc.) can be used; such mice are described in, for example, U.S. Patent No. 5,939,598.

此外，表現人類免疫球蛋白基因之替代轉染色體動物系統係此項技術中可用且可用於刺激生成(raise)本文描述之抗CD161抗體。例如，可使用攜載人類重鏈轉染色體及人類輕鏈轉染色體之小鼠(稱為「TC小鼠」)；此類小鼠描述於Tomizuka等人(2000)Proc. Natl. Acad. Sci. USA97:722-727中。此外，攜載人類重鏈及輕鏈轉殖染色體之牛已在此項技術中有所描述(Kuroiwa等人(2002)Nature Biotechnology20:889-894)且可用於刺激生成本文描述之抗CD161抗體。In addition, alternative transchromosomal animal systems expressing human immunoglobulin genes are available in this technology and can be used to stimulate the production of anti-CD161 antibodies described herein. For example, mice carrying human heavy chain transchromosomes and human light chain transchromosomes (referred to as "TC mice") can be used; such mice are described in Tomizuka et al. (2000)Proc. Natl. Acad. Sci. USA 97:722-727. In addition, cattle carrying human heavy chain and light chain transchromosomes have been described in this technology (Kuroiwa et al. (2002)Nature Biotechnology 20:889-894) and can be used to stimulate the production of anti-CD161 antibodies described herein.

此項技術中描述的用於刺激生成人類抗體(例如人類抗CD161抗體)之另外小鼠系統包括(i) Veloclmmune^®小鼠(Regeneron Pharmaceuticals, Inc.)，其中內源小鼠重鏈及輕鏈可變區已經由同源重組經人類重鏈及輕鏈可變區取代，該等人類重鏈及輕鏈可變區可操作地連接至內源小鼠恆定區，使得在該小鼠中刺激生成嵌合抗體(人類V/小鼠C)，且然後於隨後使用標準重組DNA技術轉化為完全人類抗體；及(ii) MeMo®小鼠(Merus Biopharmaceuticals, Inc.)，其中該小鼠除了單個重排的人類常見輕鏈可變區含有未重排的人類重鏈可變區。此類小鼠及其刺激生成抗體之用途描述於例如US 2012/0070861及US 2012/0073004中。Additional mouse systems described in this technology for stimulating the production of human antibodies (e.g., human anti-CD161 antibodies) include (i)^{Veloclmmune®} mice (Regeneron Pharmaceuticals, Inc.), in which the endogenous mouse heavy chain and light chain variable regions have been replaced by human heavy chain and light chain variable regions by homologous recombination, which are operably linked to endogenous mouse constant regions, so that chimeric antibodies (human V/mouse C) are stimulated in the mouse and then converted to fully human antibodies using standard recombinant DNA technology; and (ii) MeMo® mice (Merus Biopharmaceuticals, Inc.), in which the mouse contains unrearranged human heavy chain variable regions in addition to a single rearranged human common light chain variable region. Such mice and their use to stimulate the production of antibodies are described, for example, in US 2012/0070861 and US 2012/0073004.

本文描述之人類單株抗CD161抗體亦可使用用於篩選人類免疫球蛋白基因庫之噬菌體展示方法來製備。此項技術中確立此類用於分離人類抗體之噬菌體展示方法。參見例如：美國專利第5,223,409號；第5,427,908號；第5,969,108號；及第5,885,793號。The human monoclonal anti-CD161 antibodies described herein can also be prepared using phage display methods for screening human immunoglobulin gene libraries. Such phage display methods for isolating human antibodies are established in this technology. See, for example, U.S. Patent Nos. 5,223,409; 5,427,908; 5,969,108; and 5,885,793.

本文描述之人類單株抗CD161抗體亦可使用SCID小鼠來製備，該小鼠中人類免疫細胞已經重構使得在免疫時可產生人類抗體反應。此類小鼠描述於例如美國專利第5,476,996號中。The human monoclonal anti-CD161 antibodies described herein can also be prepared using SCID mice in which human immune cells have been reconstituted so that they can produce human antibody responses upon immunization. Such mice are described, for example, in U.S. Patent No. 5,476,996.

除非另有說明，否則本發明之實務將採用細胞生物學、細胞培養、分子生物學、轉殖基因生物學、微生物學、重組DNA及免疫學之習知技術，該等技術係在此項技術之技藝範圍內。此類技術在文獻中得到充分解釋。參見，例如，Sambrook等人編(1989) Molecular Cloning A Laboratory Manual (第2版；Cold Spring Harbor Laboratory Press)；Sambrook等人編 (1992) Molecular Cloning: A Laboratory Manual (Cold Springs Harbor Laboratory, NY)；D. N. Glover編, (1985) DNA Cloning, 第I卷及第II卷；Gait編(1984) Oligonucleotide Synthesis；Mullis等人，美國專利第4,683,195號；Hames及Higgins編(1984) Nucleic Acid Hybridization；Hames及Higgins編(1984) Transcription And Translation; Freshney (1987) Culture Of Animal Cells (Alan R. Liss, Inc.)；Immobilized Cells And Enzymes (IRL Press) (1986)；Perbal (1984) A Practical Guide To Molecular Cloning; the treatise, Methods In Enzymology (Academic Press, Inc., N.Y.)；Miller及Calos編(1987) Gene Transfer Vectors For Mammalian Cells (Cold Spring Harbor Laboratory)；Wu等人編, Methods In Enzymology, 第154卷及第155卷；Mayer及Walker編(1987) Immunochemical Methods In Cell And Molecular Biology (Academic Press, London)；Weir及Blackwell編(1986) Handbook Of Experimental Immunology, 第I至IV卷；Manipulating the Mouse Embryo, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1986)；Crooks, Antisense drug Technology: Principles, strategies and applications, 第2版CRC Press (2007)及Ausubel等人(1989) Current Protocols in Molecular Biology (John Wiley and Sons, Baltimore, Md.)。Unless otherwise indicated, the practice of the present invention will employ techniques of cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant DNA, and immunology, which are within the skill of the art and are fully explained in the literature. See, e.g., Sambrook et al., eds. (1989) Molecular Cloning A Laboratory Manual (2nd ed.; Cold Spring Harbor Laboratory Press); Sambrook et al., eds. (1992) Molecular Cloning: A Laboratory Manual (Cold Springs Harbor Laboratory, NY); D. N. Glover, ed. (1985) DNA Cloning, Vols. I and II; Gait, ed. (1984) Oligonucleotide Synthesis; Mullis et al., U.S. Patent No. 4,683,195; Hames and Higgins, eds. (1984) Nucleic Acid Hybridization; Hames and Higgins, eds. (1984) Transcription And Translation; Freshney (1987) Culture Of Animal Cells (Alan R. Liss, Inc.); Immobilized Cells And Enzymes (IRL Press) (1986); Perbal (1984) A Practical Guide To Molecular Cloning; the treatise, Methods In Enzymology (Academic Press, Inc., N.Y.); Miller and Calos, eds. (1987) Gene Transfer Vectors For Mammalian Cells (Cold Spring Harbor Laboratory); Wu et al., eds., Methods In Enzymology, Volumes 154 and 155; Mayer and Walker, eds. (1987) Immunochemical Methods In Cell And Molecular Biology (Academic Press, London); Weir and Blackwell (1986) Handbook Of Experimental Immunology, Volumes I to IV; Manipulating the Mouse Embryo, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1986); Crooks, Antisense drug Technology: Principles, strategies and applications, 2nd edition CRC Press (2007) and Ausubel et al. (1989) Current Protocols in Molecular Biology (John Wiley and Sons, Baltimore, Md.).

以下實例以說明方式而非藉由限制提供。實例實例1The following examples are provided by way of illustration and not by way oflimitation .

抗anti-CD161CD161單株抗體之產生Production of monoclonal antibodies

人類Ig轉殖基因小鼠用於產生抗CD161單株抗體。八隻小鼠的群組用來自AcroBiosystems之人類CD161-hFc蛋白(CD1-H5253)以及DNA免疫原表現人類及食蟹獼猴CD161 ECD域(其包括TCE-huCD161-ECD-67-225及TCE-cyCD161-ECD-67-225)免疫化。在免疫接種後第21天及第28天，使用過度表現人類及食蟹獼猴CD161之細胞系(包括CHOK1:huCD161、CHOK1:cyCD161及CHOK1:親本細胞)，藉由流動式細胞測量術檢查CD161陽性抗體效價。該八隻小鼠在第28天效價檢查之前接受四種DNA及八種蛋白質免疫接種。第28天血漿效價分析指示所有八隻免疫化小鼠中之強CD161陽性抗體效價。Human Ig transgenic mice were used to generate anti-CD161 monoclonal antibodies. Groups of eight mice were immunized with human CD161-hFc protein (CD1-H5253) from AcroBiosystems and DNA immunogens expressing human and cynomolgus macaque CD161 ECD domains, including TCE-huCD161-ECD-67-225 and TCE-cyCD161-ECD-67-225. CD161-positive antibody titers were examined by flow cytometry on days 21 and 28 after immunization using cell lines overexpressing human and cynomolgus macaque CD161, including CHOK1:huCD161, CHOK1:cyCD161, and CHOK1:parental cells. The eight mice received four DNA and eight protein immunizations prior to titer examination on day 28. Analysis of plasma titers on day 28 indicated strong CD161-positive antibody titers in all eight immunized mice.

雜交瘤產生及抗體篩選分析Hybridoma generation and antibody screening analysis

在CD161蛋白及DNA之免疫接種方案後第32天自該八隻免疫化小鼠收穫脾臟及淋巴結以產生雜交瘤。進行富含來自脾臟及淋巴結之B細胞之雜交瘤融合且接種於十四塊384孔板中。八天後，使用CHOK1:huCD161、CHOK1:cyCD161、CHOK1:P細胞(細胞混合物)進行初級多重螢光活化細胞分選(FACS)結合篩選以分離CD161陽性結合命中。對於結合篩選雜交瘤，將15 µL/孔之上清液轉移至裝納15 µL 20,000/孔細胞混合物之384孔板。在4℃下培養板30分鐘(min)且然後用FACS緩衝液洗滌2.5x。然後，用15 µL/孔之1 µg/mL AF647-山羊α人類-Fc染色該等細胞且在4℃下培養30分鐘。隨後，用FACS緩衝液洗滌2.5x細胞接著在流動式細胞儀上讀取。抗CD161參考單株抗體HP-3G10 (Biolegend；目錄號39910)用作陽性對照。Spleens and lymph nodes were harvested from the eight immunized mice on day 32 after the CD161 protein and DNA immunization regimen to generate hybridomas. Hybridomas enriched in B cells from spleen and lymph nodes were fused and seeded in fourteen 384-well plates. Eight days later, a primary multiplex fluorescence activated cell sorting (FACS) binding screen was performed using CHOK1:huCD161, CHOK1:cyCD161, CHOK1:P cells (cell mixture) to isolate CD161 positive binding hits. For binding screening hybridomas, 15 µL/well of supernatant was transferred to a 384-well plate containing 15 µL of 20,000/well cell mixture. Plates were incubated at 4°C for 30 minutes (min) and then washed 2.5x with FACS buffer. The cells were then stained with 15 µL/well of 1 µg/mL AF647-goat α-human-Fc and incubated at 4°C for 30 minutes. Subsequently, cells were washed 2.5x with FACS buffer and read on a flow cytometer. Anti-CD161 reference monoclonal antibody HP-3G10 (Biolegend; Catalog No. 39910) was used as a positive control.

自基於初級FACS之篩選，識別出134種對人類及食蟹獼猴CD161之結合劑。選擇此等以＞50K幾何平均螢光強度(gMFI)結合至HEK293T:huCD161及以＞30K gMFI結合至HEK293T:cyCD161之雜交瘤用於如圖3中所顯示之進一步驗證。然後，評估此等CD161陽性結合劑之對CD161-CLEC2D相互作用之阻斷或抑制。在134個CD161陽性結合劑中，選擇34個證實CD161與其配位體CLEC2D之相互作用之有效阻斷之雜交瘤用於次選殖。From the primary FACS-based screening, 134 binders to human and cynomolgus macaque CD161 were identified. These hybridomas that bind to HEK293T:huCD161 with >50K geometric mean fluorescent intensity (gMFI) and to HEK293T:cyCD161 with >30K gMFI were selected for further validation as shown in Figure 3. These CD161-positive binders were then evaluated for blocking or inhibition of the CD161-CLEC2D interaction. Of the 134 CD161-positive binders, 34 hybridomas that demonstrated effective blocking of the interaction of CD161 with its ligand CLEC2D were selected for secondary cloning.

篩選分析：利用Screening Analysis: UtilizationCD161CD161抗體之基於細胞之結合分析Antibody Cell-Based Binding Assays

利用CD161抗體進行基於細胞之結合分析以使用FACS評估抗CD161抗體與在CHOK1細胞表面上表現之huCD161抗原之細胞表面結合能力。表現全長膜錨定huCD161抗原之CHOK1懸浮細胞用於篩選雜交瘤抗體樣本。洗滌50,000個細胞且轉移至96孔圓底板接著添加50 µl經純化之抗體(0.001至10 µg/ml含在杜貝卡氏磷酸鹽緩衝鹽水(Dulbecco's phosphate-buffered saline，DPBS)中)且在4℃下培養30分鐘。培養後，將板以300 x g離心5分鐘。用FACS緩衝液(1% FBS含在DPBS中)進一步洗滌細胞兩次且用山羊α-小鼠IgG、Fcγ AF647 (1:2000稀釋)染色。在4℃下在黑暗中培養板20分鐘。培養後，以300 x g離心板5分鐘。用FACS緩衝液進一步洗滌細胞兩次且再次懸浮於100 µl FACS緩衝液中用於FACS分析。市售抗CD161單株Ab (HP-3G10，Biolegend)用作陽性對照及小鼠IgG1同型物(MOCP1，BioXcell)用作陰性對照。自雜交瘤產生之抗CD161抗體可與在CHOK1細胞表面上表現之huCD161抗原結合，如圖5A至5D中所示。評估代表性抗體純系(以01D17、03K18、09H07及HP-3G10 (對照)為例)在經全長huCD161構築體轉染之CHOK1上之結合，如圖5A至5C中所顯示。全劑量反應曲線顯示於圖5D中。Cell-based binding assays using CD161 antibodies were performed to assess the cell surface binding ability of anti-CD161 antibodies to huCD161 antigen expressed on the surface of CHOK1 cells using FACS. CHOK1 cell suspensions expressing full-length membrane-anchored huCD161 antigen were used to screen hybridoma antibody samples. 50,000 cells were washed and transferred to a 96-well round-bottom plate followed by the addition of 50 µl of purified antibody (0.001 to 10 µg/ml in Dulbecco's phosphate-buffered saline (DPBS)) and incubated at 4°C for 30 minutes. After incubation, the plate was centrifuged at 300 x g for 5 minutes. The cells were further washed twice with FACS buffer (1% FBS in DPBS) and stained with goat α-mouse IgG, Fcγ AF647 (1:2000 dilution). The plates were incubated at 4°C in the dark for 20 minutes. After incubation, the plates were centrifuged at 300 x g for 5 minutes. The cells were further washed twice with FACS buffer and resuspended in 100 µl FACS buffer for FACS analysis. A commercial anti-CD161 monoclonal Ab (HP-3G10, Biolegend) was used as a positive control and a mouse IgG1 isotype (MOCP1, BioXcell) was used as a negative control. Anti-CD161 antibodies produced from hybridomas can bind to huCD161 antigens expressed on the surface of CHOK1 cells, as shown in Figures 5A to 5D. Representative antibody clones (01D17, 03K18, 09H07 and HP-3G10 (control) as examples) were evaluated for binding to CHOK1 transfected with full-length huCD161 constructs, as shown in Figures 5A to 5C. The full dose response curve is shown in Figure 5D.

篩選分析：利用Screening Analysis: UtilizationCD161CD161抗體之基於Antibodies based onELISAELISA之結合分析Combination analysis

藉由ELISA評估來自雜交瘤上清液之經純化之抗CD161抗體與固定化rhCD161-hFc之結合相互作用。在4℃下，用100 µl之含在DPBS中之5 μg/mL rhCD161-hFc蛋白(R&D)塗覆高結合96孔ELISA板過夜。用ELISA洗滌緩衝液(Biolegend)洗滌該板3x，然後，用具有5%牛乳之200 µl DPBS阻斷。然後，在37℃下在溫和振盪器上以120 rpm培養板2小時(hr)，然後，用ELISA洗滌緩衝液洗滌3x。將0.0001至10 µg/ml含在100 µl分析緩衝液(1% BSA含在DPBS中)中之經純化之抗體添加至各孔且在37℃下在溫和振盪器上以120 rpm培養2小時，然後，於隨後用ELISA洗滌緩衝液洗滌3x。添加含在100 µl/孔分析緩衝液中之山羊抗小鼠偵測抗體HRP (1:3000)且在37℃下在溫和振盪器上以120 rpm培養1小時(h)然後用ELISA洗滌緩衝液洗滌5x。然後，每孔添加100 µl TMB受質且在RT下培養5分鐘。接著，添加100 µl TMB停止溶液以停止該反應。利用設定為450 nm之波長的微板讀取器讀取ELISA板之每個孔之光學密度(OD)，如圖4所示。市售抗CD161單株抗體(mAb) (HP-3G10，Biolegend)用作陽性對照及小鼠IgG1同型物(MOPC1，BioXcell)用作陰性對照。抗CD161抗體可與固定化rhCD161-hFc結合，如以顯示於圖4中之正OD值所描繪。藉由ELISA評估代表性抗CD161抗體雜交瘤純系(包括03K18、06C21、09H07、12G06及HP-3G10 (對照))與板塗覆rhCD161之結合。The binding interaction of purified anti-CD161 antibodies from hybridoma supernatants to immobilized rhCD161-hFc was assessed by ELISA. High binding 96-well ELISA plates were coated with 100 µl of 5 μg/mL rhCD161-hFc protein (R&D) in DPBS overnight at 4°C. The plates were washed 3x with ELISA wash buffer (Biolegend) and then blocked with 200 µl DPBS with 5% milk. The plates were then incubated at 37°C on a gentle shaker at 120 rpm for 2 hours (hr) and then washed 3x with ELISA wash buffer. 0.0001 to 10 µg/ml of purified antibody in 100 µl assay buffer (1% BSA in DPBS) was added to each well and incubated at 37°C on a gentle shaker at 120 rpm for 2 hours, followed by washing 3x with ELISA wash buffer. Goat anti-mouse detection antibody HRP (1:3000) in 100 µl/well assay buffer was added and incubated at 37°C on a gentle shaker at 120 rpm for 1 hour (h) followed by washing 5x with ELISA wash buffer. Then, 100 µl of TMB substrate was added per well and incubated at RT for 5 minutes. Next, 100 µl of TMB stop solution was added to stop the reaction. The optical density (OD) of each well of the ELISA plate was read using a microplate reader set to a wavelength of 450 nm, as shown in Figure 4. A commercial anti-CD161 monoclonal antibody (mAb) (HP-3G10, Biolegend) was used as a positive control and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. Anti-CD161 antibodies can bind to immobilized rhCD161-hFc, as depicted by the positive OD values shown in Figure 4. Representative anti-CD161 antibody hybridoma clones (including 03K18, 06C21, 09H07, 12G06 and HP-3G10 (control)) were evaluated for binding to plate-coated rhCD161 by ELISA.

篩選分析：基於細胞之靶及配位體阻斷分析Screening Assays: Cell-based Target and Ligand Blockade Assays

如Kamishikiryo等人(2011)所報告，CD161之CLEC2D親和力為48 μM之K_D。評估使用抗體阻斷CD161-CLEC2D相互作用。使用huCLEC2D可溶性蛋白及表現huCD161之CHOK1細胞評估抗CD161陽性雜交瘤命中阻斷CD161-CLEC2D相互作用之能力。基於CD161-CLEC2D之間的較低親和力，需要產生CLEC2D蛋白之多聚物複合物以增加價度。As reported by Kamishikiryo et al. (2011), the affinity of CD161 for CLEC2D is a_KD of 48 μM. The use of antibodies to block the CD161-CLEC2D interaction was evaluated. The ability of anti-CD161-positive hybridoma hits to block the CD161-CLEC2D interaction was evaluated using huCLEC2D soluble protein and CHOK1 cells expressing huCD161. Based on the low affinity between CD161-CLEC2D, it is necessary to generate multimeric complexes of CLEC2D protein to increase valency.

為了產生CLEC2D多聚物，將生物素化蛋白A (ThermoFisher)及hCLEC2D-hFc蛋白(Kactus)以1:20比混合且在4℃下在黑暗中培養5分鐘。將PBS添加至該混合物且在4℃下在黑暗中培養30分鐘。培養後，添加(在DPBS中1:1000稀釋)鏈黴親和素-APC珠粒(Biolegend)且在4℃下在黑暗中培養30分鐘。添加經純化之人類IgG (ThermoFisher)用於Fc飽和。To generate CLEC2D multimers, biotinylated protein A (ThermoFisher) and hCLEC2D-hFc protein (Kactus) were mixed at a 1:20 ratio and incubated at 4°C in the dark for 5 minutes. PBS was added to the mixture and incubated at 4°C in the dark for 30 minutes. After incubation, streptavidin-APC beads (Biolegend) were added (diluted 1:1000 in DPBS) and incubated at 4°C in the dark for 30 minutes. Purified human IgG (ThermoFisher) was added for Fc saturation.

表現huCD161抗原之CHOK1細胞用於篩選抗CD161抗體樣本。洗滌50,000至100,000個細胞且轉移至96孔圓底板。將預製備的hCLEC2D多聚物複合物(20 ul)加上50 µl之經純化之CD161抗體樣本添加至CD161表現細胞且在4℃下在黑暗中培養30分鐘。培養後，以300 x g離心板5分鐘。用FACS緩衝液進一步洗滌細胞兩次且再次懸浮於100 µl FACS緩衝液中用於FACS分析。市售抗CD161 mAb (HP-3G10，Biolegend)用作陽性對照及小鼠IgG1同型物(MOPC1，BioXcell)用作陰性對照。使用以下公式計算CD161-CLEC2D相互作用之阻斷百分比：阻斷的% = (1 – 具有抗體下結合至CHOK1-CD161之CLEC2D多聚物%/不具有抗體下結合至CHOK1-CD161之CLEC2D多聚物%) x 100%。如圖6中所示，hCLEC2D多聚物有效地結合至表現huCD161之CHOK1。10 ug/ml之參考抗CD161單株Ab (HP-3G10)導致hCLEC2D多聚物之完全阻斷(圖6)。能夠阻斷CLEC2D多聚物與在細胞表面上表現之huCD161之間的相互作用之抗CD161抗體之阻斷百分比顯示於圖7中。藉由FACS及結合至表現CD161之CHOK1之CLEC2D多聚物來評估來自所選雜交瘤之抗CD161抗體。基於hCLEC2D多聚物與hCD161之間的相互作用之阻斷來排序阻斷之百分比。CHOK1 cells expressing huCD161 antigen were used to screen anti-CD161 antibody samples. 50,000 to 100,000 cells were washed and transferred to 96-well round-bottom plates. Pre-prepared hCLEC2D multimer complex (20 ul) plus 50 µl of purified CD161 antibody sample were added to CD161 expressing cells and incubated at 4°C in the dark for 30 minutes. After incubation, the plate was centrifuged at 300 x g for 5 minutes. Cells were further washed twice with FACS buffer and resuspended in 100 µl FACS buffer for FACS analysis. A commercial anti-CD161 mAb (HP-3G10, Biolegend) was used as a positive control and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. The percentage of blockade of the CD161-CLEC2D interaction was calculated using the following formula: % Blocked = (1 – % CLEC2D multimers bound to CHOK1-CD161 with antibody/% CLEC2D multimers bound to CHOK1-CD161 without antibody) x 100%. As shown in FIG6 , hCLEC2D multimers bind efficiently to CHOK1 expressing huCD161. 10 ug/ml of the reference anti-CD161 monoclonal Ab (HP-3G10) resulted in complete blockade of hCLEC2D multimers ( FIG6 ). The percentage of blocking of anti-CD161 antibodies that were able to block the interaction between CLEC2D multimers and huCD161 expressed on the cell surface is shown in Figure 7. Anti-CD161 antibodies from selected hybridomas were evaluated by FACS and bound to CLEC2D multimers of CHOK1 expressing CD161. The percentage of blocking was ranked based on the blocking of the interaction between hCLEC2D multimers and hCD161.

篩選分析：基於Screening analysis: Based onELISAELISA之OfCD161:CLEC2DCD161:CLEC2D阻斷分析Blockage Analysis

藉由ELISA評估抗CD161抗體阻斷rhCLEC2D與固定化rhCD161-hFc之間的相互作用之能力。在4℃下，將高結合96孔ELISA板塗覆含在DPBS中之100 µl rhCD161-hFc (5 μg/ml)蛋白(R&D)過夜。用ELISA洗滌緩衝液(Biolegend)洗滌板3x然後用用具有5%牛乳之200 µl DPBS阻斷且在37℃下在溫和振盪器上以120 rpm培養2小時接著用ELISA洗滌緩衝液洗滌3x。將含在100 µl分析緩衝液(1% BSA含在DPBS中)中之生物素化10 μg /ml CLEC2D-hFc (Kactus)及0.03至66.7 nM之經純化之抗體添加至各孔且在37℃下在溫和振盪器上以120 rpm培養2小時接著於隨後用ELISA洗滌緩衝液洗滌3x。將含在100 µl分析緩衝液中之抗生物素蛋白HRP (1:1000)添加至該板之各孔且在37℃下在溫和振盪器上以120 rpm培養1小時接著用ELISA洗滌緩衝液洗滌5x。將體積為100 µl之TMB受質添加至各孔且在37℃下培養30分鐘接著添加100 µl TMB停止溶液以停止該反應。利用設定為450 nm之波長的微板讀取器讀取該ELISA板之每個孔之光學密度(OD)。市售抗CD161 mAb (HP-3G10，Biolegend)用作陽性對照及小鼠IgG1同型物(MOPC1，BioXcell)用作陰性對照。如圖8中所繪示，抗CD161抗體可阻斷rhCLEC2D與固定化rhCD161-hFc之間的相互作用。評估代表性抗CD161雜交瘤純系(包括01D17、03K18、12G06及HP-3G10 (陽性對照))之與CLEC2D-CD161接觸位點之競爭及對該相互作用之破壞。 實例2. 抗CD161單株抗體之功能評估The ability of anti-CD161 antibodies to block the interaction between rhCLEC2D and immobilized rhCD161-hFc was assessed by ELISA. High binding 96-well ELISA plates were coated with 100 µl rhCD161-hFc (5 μg/ml) protein (R&D) in DPBS overnight at 4°C. Plates were washed 3x with ELISA wash buffer (Biolegend) then blocked with 200 µl DPBS with 5% milk and incubated at 37°C on a gentle shaker at 120 rpm for 2 hours followed by washing 3x with ELISA wash buffer. Biotinylated 10 μg/ml CLEC2D-hFc (Kactus) and 0.03 to 66.7 nM purified antibodies in 100 μl assay buffer (1% BSA in DPBS) were added to each well and incubated at 37°C on a gentle shaker at 120 rpm for 2 hours followed by washing 3x with ELISA wash buffer. Avidin HRP (1:1000) in 100 μl assay buffer was added to each well of the plate and incubated at 37°C on a gentle shaker at 120 rpm for 1 hour followed by washing 5x with ELISA wash buffer. A volume of 100 µl of TMB substrate was added to each well and incubated at 37°C for 30 minutes followed by the addition of 100 µl of TMB stop solution to stop the reaction. The optical density (OD) of each well of the ELISA plate was read using a microplate reader set to a wavelength of 450 nm. A commercial anti-CD161 mAb (HP-3G10, Biolegend) was used as a positive control and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. As shown in FIG8 , the anti-CD161 antibody blocked the interaction between rhCLEC2D and immobilized rhCD161-hFc. Representative anti-CD161 hybridoma clones (including 01D17, 03K18, 12G06 and HP-3G10 (positive control)) were evaluated for competition with the CLEC2D-CD161 contact site and disruption of the interaction.Example 2. Functional evaluation of anti-CD161 monoclonal antibodies

利用金黃色葡萄球菌腸毒素Using Staphylococcus aureus enterotoxinBB刺激之StimulationTT細胞活性評估Cell viability assessment

在37℃下，在完全細胞培養基(RPMI + 10% FBS)中含有或不含測試抗CD161抗體下用10 ng/ml金黃色葡萄球菌腸毒素B (SEB)刺激來自健康人類供體之周邊血液單核細胞(PBMC) 24小時，接著測定細胞培養上清液中之IL-2細胞激素。市售抗CD161單株抗體(mAb) (HP-3G10，Biolegend)用作陽性對照及小鼠IgG1同型物(MOPC1，BioXcell)用作陰性對照。使用以下公式計算IL-2細胞激素之百分比增加：增加的% = (具有抗體之樣本之細胞激素濃度 / 不具有抗體之樣本之細胞激素濃度) x 100% - 100%。Peripheral blood mononuclear cells (PBMCs) from healthy human donors were stimulated with 10 ng/ml Staphylococcus aureus enterotoxin B (SEB) in complete cell culture medium (RPMI + 10% FBS) with or without test anti-CD161 antibodies at 37°C for 24 hours, and IL-2 cytokines were measured in cell culture supernatants. A commercial anti-CD161 monoclonal antibody (mAb) (HP-3G10, Biolegend) was used as a positive control and a mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. The percent increase in IL-2 cytokine was calculated using the following formula: % increase = (cytokine concentration of sample with antibody / cytokine concentration of sample without antibody) x 100% - 100%.

圖9中顯示可藉由阻斷CD161-CLEC2D相互作用來增強T細胞功能(藉由IL-2細胞激素釋放測得)之抗CD161抗體，一種自一名供體之PBMC上清液之IL-2細胞激素表現之代表性ELISA分析。相較於SEB刺激之PBMC中不含添加之測試抗體，基於IL-2釋放程度對抗CD161抗體進行排序作為IL-2的增加百分比(圖9)。A representative ELISA analysis of IL-2 cytokine expression in PBMC supernatant from one donor is shown in Figure 9 for anti-CD161 antibodies that can enhance T cell function (as measured by IL-2 cytokine release) by blocking CD161-CLEC2D interaction. Anti-CD161 antibodies were ranked based on the degree of IL-2 release as a percentage increase in IL-2 compared to SEB-stimulated PBMCs without added test antibody (Figure 9).

藉由ByTT細胞接合劑介導之Cell Adaptor-mediatedTT細胞活性Cell activity

ELISA分析用於分析PBMC上清液中之IL-2細胞激素含量，且基於IL-2釋放評估抗CD161抗體且進行排序。對於此分析，洗滌Raji細胞且以30,000個細胞/孔轉移至96孔圓底板接著在37℃下用100 µg/ml CD3xCD19 T細胞接合劑(InvivoGen)預培養30分鐘。將一定量之90,000個表現全長膜錨定huCD161抗原之Jurkat細胞連同經純化之抗CD161抗體及人類Fc阻斷劑(1:1000)添加至該Raji/T細胞接合劑板。在37℃下培養板48小時，接著測定細胞培養上清液中之細胞激素。市售抗CD161 mAb (HP-3G10，Biolegend)用作陽性對照及小鼠IgG1同型物(MOPC1，BioXcell)用作陰性對照。如圖10中所繪示，藉由阻斷hCD161-hCLEC2D相互作用，抗-CD161抗體可增強表現huCD161之Jurkat細胞功能，藉由IL-2釋放測定。 實例3. 抗CD161單株抗體在抑制CD161-CLEC2D相互作用時增強T細胞之活化ELISA analysis was used to analyze IL-2 cytokine levels in PBMC supernatants, and anti-CD161 antibodies were evaluated and ranked based on IL-2 release. For this analysis, Raji cells were washed and transferred to 96-well round-bottom plates at 30,000 cells/well and then pre-incubated with 100 µg/ml CD3xCD19 T cell binder (InvivoGen) for 30 minutes at 37°C. An amount of 90,000 Jurkat cells expressing full-length membrane-anchored huCD161 antigen was added to the Raji/T cell binder plate along with purified anti-CD161 antibodies and human Fc blocker (1:1000). Plates were incubated at 37°C for 48 hours, followed by assay of cytokines in cell culture supernatants. Commercially available anti-CD161 mAb (HP-3G10, Biolegend) was used as a positive control and mouse IgG1 isotype (MOPC1, BioXcell) was used as a negative control. As shown in Figure 10, by blocking the hCD161-hCLEC2D interaction, anti-CD161 antibodies can enhance the function of Jurkat cells expressing huCD161, as measured by IL-2 release.Example 3. Anti-CD161 monoclonal antibodies enhance T cell activation when inhibiting CD161-CLEC2D interaction

分析抗CD161單株抗體(mAb)在MART-1肽(HLA-A2限制)的存在下救援藉由過度表現hCLEC2D之MeWo細胞(HLA-A2+)活化MART-1 TCR+hCD161+ Jurkat細胞之能力。The ability of anti-CD161 monoclonal antibodies (mAbs) to rescue MART-1 TCR+hCD161+ Jurkat cells activated by MeWo cells (HLA-A2+) overexpressing hCLEC2D in the presence of MART-1 peptide (HLA-A2 restricted) was analyzed.

方法method

將模擬-GFP及過度表現hCLEC2D-GFP之MeWo細胞系(HLA；A*0201，其係為肽呈現所需)以20K/孔培養於FB 96孔板中過夜(參見圖11)。第二天，以300 g/5分鐘離心板，在不觸及黏附MeWo細胞下小心移除培養基，且然後添加0.1 ug/孔之在RPMI完全培養基中製備之MART-1肽，接著培養細胞30分鐘/37℃。在培養期間，製備圓底板50 µL之具有過度表現人類CD161之細胞(100K/孔)之Jurkat-MART-1 TCR，且然後以自0.005至10 µg/mL之最終濃度添加50 µL抗CD161 mAb。在肽板培養之後，在96孔板中在具有肽FB之MeWo細胞的頂部添加具有抗CD161 mAb 100 µL之Jurkat-MART1-CD161 Oe細胞系且在37℃下培養24小時。第二天，培養後，以500 g離心板5分鐘，且使用Biolegend套組(目錄號431816批次：B352552；按照製造商說明書)，收集180 µL上清液用於IL-2 ELISA。Mock-GFP and MeWo cell lines overexpressing hCLEC2D-GFP (HLA; A*0201, which is required for peptide presentation) were cultured in FB 96-well plates at 20K/well overnight (see Figure 11). The next day, the plates were centrifuged at 300 g/5 minutes, the medium was carefully removed without touching the adherent MeWo cells, and then 0.1 ug/well of MART-1 peptide prepared in RPMI complete medium was added, followed by culturing the cells for 30 minutes/37°C. During the culture period, round bottom plates 50 µL of Jurkat-MART-1 TCR with cells overexpressing human CD161 (100K/well) were prepared and then 50 µL of anti-CD161 mAb was added at final concentrations ranging from 0.005 to 10 µg/mL. After peptide plate incubation, Jurkat-MART1-CD161 Oe cell line with anti-CD161 mAb 100 µL was added on top of MeWo cells with peptide FB in 96-well plates and cultured at 37°C for 24 hours. On the second day, after incubation, the plates were centrifuged at 500 g for 5 min and 180 µL of supernatant was collected for IL-2 ELISA using the Biolegend kit (Cat. No. 431816 Batch: B352552; according to the manufacturer's instructions).

結果result

每種抗CD161 mAb引發TCR活化相較於同型物對照之劑量依賴性增加，藉由IL-2分泌測定(圖12)。此等結果指示，TCR活化受到人類CLEC2D的影響，且其可利用抗CD161抗體救援，此阻斷CD161受體(Jurkat)與其在MeWo細胞上表現之配位體CLEC2D之間的相互作用。如所期，對照hIgG1 LALA-PG抗體未增加進一步的TCR活化。Each anti-CD161 mAb elicited a dose-dependent increase in TCR activation relative to the isotype control, as measured by IL-2 secretion (Figure 12). These results indicate that TCR activation is affected by human CLEC2D and that it can be rescued using anti-CD161 antibodies, which block the interaction between the CD161 receptor (Jurkat) and its ligand CLEC2D expressed on MeWo cells. As expected, the control hIgG1 LALA-PG antibody did not increase further TCR activation.

因此，抗CD161抗體抑制CD161與CLEC2D之間的負相互作用且增強T細胞活化，藉由IL-2釋放測定。 實例4. 抗CD161單株抗體結合至TALL-104細胞且救援藉由TALL-104細胞對過度表現hCLEC2D之靶細胞之細胞溶解Thus, anti-CD161 antibodies inhibit the negative interaction between CD161 and CLEC2D and enhance T cell activation as measured by IL-2 release.Example 4. Anti-CD161 monoclonal antibodies bind to TALL-104 cells and rescue cytolysis of target cells overexpressing hCLEC2D by TALL-104 cells

分析抗CD161 mAb結合TALL-104效應細胞且救援藉由CD161+ TALL-104作為效應細胞(CD3/TCR+ CD4- CD8+ CD56+ CD16-)對過度表現hCLEC2D之(OE) PC3靶細胞之細胞溶解之能力。TALL-104效應細胞係臨床相關MHC非限制性人類細胞毒性T細胞系。The ability of anti-CD161 mAb to bind TALL-104 effector cells and rescue cytolysis of hCLEC2D-overexpressing (OE) PC3 target cells by CD161+ TALL-104 as effector cells (CD3/TCR+ CD4- CD8+ CD56+ CD16-) was analyzed. TALL-104 effector cells are a clinically relevant MHC-unrestricted human cytotoxic T cell line.

TALL-104TALL-104結合Combine

洗滌TALL-104細胞且計數，然後以100,000個細胞/孔放置於96孔圓底板中。將板以500 g離心5分鐘，且丟棄上清液。接下來，添加在FACS緩衝液中製備的50 µL抗CD161 mAb，將樣本混合，且然後在冰上培養30分鐘。培養後，用100 µL FACS緩衝液(DPBS+1%FBS)洗滌板，以500 g離心5分鐘，且丟棄上清液。然後，將板與50 µL山羊抗人類IgG (H+L)交叉吸附二級抗體、具有zombie aqua之ALEXA FLUOR™ Plus 647及1:2000稀釋的活死染料在冰上培養20分鐘。培養後，用100 µL FACS緩衝液(DPBS + 1% FBS)洗滌板。然後，將板以500 g離心5分鐘。然後，丟棄上清液且將細胞沉澱再懸浮於100 µL/孔FACS緩衝液中接著進行流動式細胞測量儀之採集。Wash TALL-104 cells and count, then plate at 100,000 cells/well in a 96-well round-bottom plate. Centrifuge the plate at 500 g for 5 minutes, and discard the supernatant. Next, add 50 µL of anti-CD161 mAb prepared in FACS buffer, mix the samples, and then incubate on ice for 30 minutes. After incubation, wash the plate with 100 µL FACS buffer (DPBS+1%FBS), centrifuge at 500 g for 5 minutes, and discard the supernatant. The plate was then incubated on ice for 20 minutes with 50 µL of goat anti-human IgG (H+L) cross-adsorbed secondary antibody, ALEXA FLUOR™ Plus 647 with zombie aqua, and 1:2000 dilution of live-dead dye. After incubation, the plate was washed with 100 µL of FACS buffer (DPBS + 1% FBS). The plate was then centrifuged at 500 g for 5 minutes. The supernatant was then discarded and the cell pellet was resuspended in 100 µL/well FACS buffer followed by flow cytometry acquisition.

所測試的抗CD161 mAbs中之各者能夠以劑量依賴性方式結合TALL-104細胞(圖13)。Each of the tested anti-CD161 mAbs was able to bind to TALL-104 cells in a dose-dependent manner ( FIG. 13 ).

救援RescueTALL-104TALL-104效應功能Effect function

基於xCELLigence之細胞毒性分析用於評估抗CD161 mAb救援藉由CD161+ TALL-104效應細胞(CD3/TCR+ CD4- CD8+ CD56+ CD16-)對過度表現hCLEC2D之PC3靶細胞之細胞溶解之能力(圖14A)。將CD161+ TALL-104人類殺手CD8+ T細胞與CLEC2D+腫瘤細胞(PC3-CLEC2D OE)培養。相較於WT PC3細胞(CLEC2D陰性)，PC3腫瘤細胞上CLEC2D之過度表現會抑制TALL-104介導之殺死(圖14B至圖14C)。測試上文顯示阻斷CD161-CLEC2D相互作用之抗CD161 mAb之增強TALL-104介導之殺死PC3-CLEC2D OE之能力。xCELLigence-based cytotoxicity assays were used to evaluate the ability of anti-CD161 mAbs to rescue cytolysis of PC3 target cells overexpressing hCLEC2D by CD161+ TALL-104 effector cells (CD3/TCR+ CD4- CD8+ CD56+ CD16-) (FIG. 14A). CD161+ TALL-104 human killer CD8+ T cells were cultured with CLEC2D+ tumor cells (PC3-CLEC2D OE). Overexpression of CLEC2D on PC3 tumor cells inhibited TALL-104-mediated killing compared to WT PC3 cells (CLEC2D negative) (FIG. 14B-C). The anti-CD161 mAbs shown above to block CD161-CLEC2D interaction were tested for their ability to enhance TALL-104-mediated killing of PC3-CLEC2D OE.

步驟Steps11：黏附靶細胞：Adhesion to target cells((亦即that is，,腫瘤細胞Tumor cells– PC3)– PC3)

在第-1天，將模擬/OE PC-3細胞胰蛋白酶化且進行計數。然後，將細胞以10K個細胞/孔接種於96孔E板中過夜。On day -1, mock/OE PC-3 cells were trypsinized and counted. Then, cells were seeded at 10K cells/well in 96-well E plates overnight.

步驟Steps22：:TALL-104TALL-104添加作為效應細胞Add as effector cells((亦即免疫細胞Immune cells))

第二天，計數TALL-104細胞且離心。然後，將TALL-104細胞再懸浮於培養基中且用抗-CD161 mAb或同型物IgG1 LALA-PG對照處理30分鐘/37℃。培養後，將TALL-104細胞與PC3細胞以1:1比在E板中接觸且在RT下培養30分鐘以促進PC3靶細胞的頂部上TALL-104細胞之均勻分佈。然後，將該等板放置回至位於培養箱內部的xCELLigence儀器中，且在37℃下採集數據20小時。The next day, TALL-104 cells were counted and centrifuged. TALL-104 cells were then resuspended in medium and treated with anti-CD161 mAb or isotype IgG1 LALA-PG control for 30 minutes/37°C. After incubation, TALL-104 cells were contacted with PC3 cells in an E plate at a 1:1 ratio and incubated at RT for 30 minutes to promote even distribution of TALL-104 cells on top of PC3 target cells. The plates were then placed back into the xCELLigence instrument inside the incubator and data were collected for 20 hours at 37°C.

步驟Steps33：若效應細胞誘導靶黏附腫瘤細胞之破壞，則可敏感且精確地偵測此細胞溶解活性：If effector cells induce the destruction of target adherent tumor cells, this cytolytic activity can be detected sensitively and accurately

xCELLigence RTCA無標籤技術使用嵌入於專有E板中之金電極之阻抗變化來計數細胞。在此xCELLigence分析中，將hCLEC2D OE PC3腫瘤細胞黏附至嵌入於微量滴定板(E板)底部的叉指式金微電極之表面。靶細胞與金感測器之相互作用產生阻抗信號，其反映靶細胞的數量、尺寸及黏附強度。效應TALL-104細胞不是天然黏附的事實簡化此系統，因為其產生可容易地減去的低阻抗信號。然而，當TALL-104經由細胞溶解攻擊且殺死PC3細胞時，靶PC3細胞之存活率藉由阻抗之即時變化來反映，從而提供殺死之動力學評估。TALL-104的細胞溶解活性導致黏附細胞集合(round up)及分離，從而導致較低的CI值。The xCELLigence RTCA label-free technology uses changes in impedance of a gold electrode embedded in a proprietary E-plate to count cells. In this xCELLigence assay, hCLEC2D OE PC3 tumor cells are adhered to the surface of an interdigitated gold microelectrode embedded in the bottom of a microtiter plate (E-plate). The interaction of the target cells with the gold sensor produces an impedance signal that reflects the number, size, and adhesion strength of the target cells. The fact that effector TALL-104 cells are not naturally adherent simplifies this system because they produce a low impedance signal that can be easily subtracted. However, when TALL-104 attacks and kills PC3 cells by cytolysis, the viability of the target PC3 cells is reflected by real-time changes in impedance, providing a kinetic assessment of the killing. The cytolytic activity of TALL-104 resulted in the round up and dissociation of adherent cells, leading to lower CI values.

RTCA系統使用細胞阻抗讀數以監測細胞數量、細胞尺寸及細胞-受質黏附強度之即時變化作為稱為細胞指數(CI)的單一參數來反映靶PC3細胞之存活率。為了證實此方法作為TALL-104細胞介導之靶PC3細胞之細胞溶解之效價分析之有效性。首先，將經工程化為模擬作為對照(模擬-PC3)或過度表現人類CLEC2D之細胞(CLEC2D OE PC3)之PC3前列腺癌細胞於E板中培養且使用xCELLigence測量其增殖。在接種PC3靶細胞的次日，將TALL-104效應細胞以1:1之E:T比添加至該等孔，且監測靶PC3細胞的存活率。用單獨效應細胞生長培養基處理之PC3細胞充當陰性對照。將TALL-104細胞添加至PC3細胞導致CI之立即且時間依賴性降低。由於細胞溶解，PC3信號在添加效應細胞約24小時後下降。The RTCA system uses cell impedance readings to monitor real-time changes in cell number, cell size, and cell-substrate adhesion strength as a single parameter called the cell index (CI) to reflect the viability of target PC3 cells. To demonstrate the effectiveness of this method as a titer assay for TALL-104 cell-mediated cytolysis of target PC3 cells, PC3 prostate cancer cells engineered to mimic as a control (Mock-PC3) or cells overexpressing human CLEC2D (CLEC2D OE PC3) were first cultured in E plates and their proliferation was measured using xCELLigence. The day after inoculation of PC3 target cells, TALL-104 effector cells were added to the wells at a 1:1 E:T ratio and the survival of the target PC3 cells was monitored. PC3 cells treated with effector cell growth medium alone served as a negative control. Addition of TALL-104 cells to PC3 cells resulted in an immediate and time-dependent reduction in CI. PC3 signal decreased approximately 24 hours after addition of effector cells due to cell lysis.

每個抗CD161 mAb僅在人類CLEC2D OE-PC3細胞中顯示細胞溶解之劑量依賴性增加(圖15)，這指示基於TALL-104之PC3殺死受到人類CLEC2D的影響且可利用阻斷CD161受體(TALL-104)與其在PC3細胞上表現之配位體CLEC2D之間的相互作用之抗CD161抗體救援。 實例5. 抗CD161 mAb增加NK細胞毒性Each anti-CD161 mAb showed a dose-dependent increase in cytolysis only in human CLEC2D OE-PC3 cells (Figure 15), indicating that TALL-104-based PC3 killing is affected by human CLEC2D and can be rescued by anti-CD161 antibodies that block the interaction between the CD161 receptor (TALL-104) and its ligand CLEC2D expressed on PC3 cells.Example 5. Anti-CD161 mAbs increase NK cell cytotoxicity

分析抗CD161 mAb於初級人類NK細胞細胞毒性上之效應。將人類NK細胞用200 U/mL IL-2活化過夜，然後與細胞示蹤紫(cell trace violet，CTV)標記之PC-3 CLEC2D-OE (過度表現CLEC2D)或Raji細胞在Fc阻斷加上1 ug/mL抗CD161 mAb或hIgG1-LALAPG對照抗體之存在下共培養18小時。藉由CTV+群體內的腫瘤細胞存活率(Zombie+)來測量於腫瘤細胞上之細胞毒性。The effect of anti-CD161 mAb on primary human NK cell cytotoxicity was analyzed. Human NK cells were activated overnight with 200 U/mL IL-2 and then co-cultured with PC-3 CLEC2D-OE (overexpressing CLEC2D) or Raji cells labeled with cell trace violet (CTV) for 18 hours in the presence of Fc blockade plus 1 ug/mL anti-CD161 mAb or hIgG1-LALAPG control antibody. Cytotoxicity on tumor cells was measured by tumor cell survival within the CTV+ population (Zombie+).

相較於同型物對照，與抗CD161 mAb接觸導致對PC-3 CLEC2D-OE (圖16A)或Raji (圖16B)細胞之增加之NK細胞毒性，經由相對腫瘤殺死能力所評估。相對腫瘤殺死能力藉由在抗CD161 mAb之存在下將腫瘤細胞死亡(%)標準化至同型物來計算得。數據代表每個抗CD161純系之跨4或3個獨立分析之PC-3 CLEC2D-OE的九名健康供體(圖16A)及Raji的七名健康供體(圖16B)。 實例6. 抗CD161 mAb增強NK細胞對腫瘤生長之抑制Exposure to anti-CD161 mAb resulted in increased NK cytotoxicity against PC-3 CLEC2D-OE (Figure 16A) or Raji (Figure 16B) cells compared to isotype controls, as assessed by relative tumor killing capacity. Relative tumor killing capacity was calculated by normalizing tumor cell death (%) to isotype in the presence of anti-CD161 mAb. Data represent nine healthy donors for PC-3 CLEC2D-OE (Figure 16A) and seven healthy donors for Raji (Figure 16B) across 4 or 3 independent analyses for each anti-CD161 clone.Example 6. Anti-CD161 mAb enhances NK cell inhibition of tumor growth

健康初始雌性小鼠(6至8週；NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ NSG；Jackson Laboratory)經皮下注射2 x 10⁶個PC3-CLEC2D-OE細胞。當腫瘤尺寸達到約70 mm³(第4天)時，將小鼠隨機分組至治療組(n=4)。前兩劑的抗CD161抗體12G06 (10 mg/kg)經靜脈內注射，且後六劑(15 mg/kg)經腹膜內注射。治療於第4天開始且每3天重複。Healthy naive female mice (6 to 8 weeks; NOD.Cg-Prkdc^scid Il2rg^tm1Wjl /SzJ NSG; Jackson Laboratory) were injected subcutaneously with 2 x 10⁶ PC3-CLEC2D-OE cells. When the tumor size reached approximately 70 mm³ (day 4), the mice were randomized into treatment groups (n=4). The first two doses of anti-CD161 antibody 12G06 (10 mg/kg) were injected intravenously, and the last six doses (15 mg/kg) were injected intraperitoneally. Treatment started on day 4 and repeated every 3 days.

所有小鼠在第4天及第10天接受兩次5 x 10⁶個人類NK細胞的瘤內投與。自冷凍人類PBMC新鮮分離NK細胞且在注射前一晚與重組人類IL-2 100U/mL在NK MACS培養基(Miltenyi Biotech)中培養。All mice received two intratumoral administrations of 5 x¹⁰⁶ human NK cells on days 4 and 10. NK cells were freshly isolated from frozen human PBMCs and cultured in NK MACS medium (Miltenyi Biotech) with recombinant human IL-2 100 U/mL the night before injection.

利用測徑規在二維度測量腫瘤生長，且使用公式(寬度²x 長度)/2計算腫瘤體積(mm³)。Tumor growth was measured in two dimensions using calipers, and tumor volume (mm³ ) was calculated using the formula (width² x length)/2.

相較於媒劑對照組，在異種移植模型中，在NK細胞存在下，12G06抗體之投與顯著增強腫瘤生長抑制(圖17)。將利用另外抗CD161抗體(例如01C07、09O13、12H24、06M03 S及/或06M03 L)重複此等實驗。 實例7. 抗CD161 mAb增強藉由共同植入的PBMC對腫瘤生長之抑制Administration of 12G06 antibody significantly enhanced tumor growth inhibition in the presence of NK cells in a xenograft model compared to vehicle control (Figure 17). These experiments will be repeated using additional anti-CD161 antibodies (e.g., 01C07, 09O13, 12H24, 06M03 S and/or 06M03 L).Example 7. Anti-CD161 mAb enhances inhibition of tumor growth by co-implanted PBMCs

健康小鼠(NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ NSG；Jackson Laboratory)經皮下注射PBMC及U-87 MG (神經膠質母細胞瘤)人類腫瘤細胞之1:4混合物之4.5 x 10⁶個細胞至右腹側。在植入後1小時經腹膜內投與抗CD161抗體12G06，且小鼠每3天接受總共八劑的20 mg/kg 12G06。Healthy mice (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl /SzJ NSG; Jackson Laboratory) were injected subcutaneously into the right flank with 4.5 x 10⁶ cells of a 1:4 mixture of PBMC and U-87 MG (neuroglioblastoma) human tumor cells. Anti-CD161 antibody 12G06 was administered intraperitoneally 1 hour after implantation, and mice received a total of eight doses of 20 mg/kg 12G06 every 3 days.

利用測徑規在二維度測量腫瘤生長，且使用公式(寬度²x 長度)/2計算腫瘤體積(mm³)。Tumor growth was measured in two dimensions using calipers, and tumor volume (mm³ ) was calculated using the formula (width² x length)/2.

相較於媒劑對照組，在異種移植模型中，在人類PBMC存在下，12G06抗體之投與顯著增強腫瘤生長抑制(圖18)。將利用另外抗CD161抗體(例如01C07、09O13、12H24、06M03 S及/或06M03 L)重複此等實驗。 實例8. 抗CD161抗體於人類癌症患者中之活體內分析(預示)Administration of 12G06 antibody significantly enhanced tumor growth inhibition in the presence of human PBMCs in a xenograft model compared to vehicle control (Figure 18). These experiments will be repeated using additional anti-CD161 antibodies (e.g., 01C07, 09O13, 12H24, 06M03 S and/or 06M03 L).Example 8. In vivo analysis of anti-CD161 antibodies in human cancer patients (predictive)

將在人類癌症患者中測試抗CD161抗體之安全性及功效。將對患者投與一劑量之12G06、01C07、09O13、12H24、06M03 S及/或06M03 L抗體。將監測患者之任何不良事件，且測定腫瘤負擔、無疾病進展存活期及總存活期。The safety and efficacy of anti-CD161 antibodies will be tested in human cancer patients. Patients will be administered a dose of 12G06, 01C07, 09O13, 12H24, 06M03 S and/or 06M03 L antibodies. Patients will be monitored for any adverse events, and tumor burden, disease progression-free survival, and overall survival will be determined.

應明瞭，[實施方式]部分而非[發明內容]及[摘要]部分意欲用於解釋申請專利範圍。[發明內容]及[摘要]部分可闡述發明人所設想的本發明之一或多個而非所有示例性態樣，且因此，不欲以任何方式限制本發明及隨附申請專利範圍。It should be understood that the [Implementation] section, rather than the [Contents of the Invention] and [Abstract] sections, is intended to be used to interpret the scope of the patent application. The [Contents of the Invention] and [Abstract] sections may describe one or more but not all exemplary aspects of the invention contemplated by the inventor, and therefore, are not intended to limit the scope of the invention and the accompanying patent application in any way.

已於上文藉助於繪示指定功能及其關係之實施方案之功能構建塊描述本發明。為了方便描述，在本文中已任意定義此等功能構建塊之邊界。可定義替代邊界，只要適當地進行指定功能及其關係即可。The present invention has been described above with the aid of functional building blocks that illustrate implementations of specified functions and relationships. For ease of description, the boundaries of these functional building blocks have been arbitrarily defined herein. Alternative boundaries may be defined as long as the specified functions and relationships are appropriately performed.

特定態樣之前述描述將如此充分地揭示本發明之一般性質以致於其他人可在不脫離本發明之一般概念下藉由應用此項技術的技藝內的知識容易地修改及/或適用於此類特定態樣的各種應用而無須過度實驗。因此，基於本文呈現的教示及指導，此類調適及修改意欲在所揭示的態樣之等效物之含義及範圍內。應理解，本文的用語或術語係出於描述而非限制之目的，使得本說明書之術語或用語意欲由熟練技術者根據教示及指導來解釋。The foregoing description of specific aspects will fully disclose the general nature of the invention so that others can easily modify and/or adapt to various applications of such specific aspects without undue experimentation by applying the knowledge in the art of this technology without departing from the general concept of the invention. Therefore, based on the teachings and guidance presented herein, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed aspects. It should be understood that the terms or terms herein are for the purpose of description rather than limitation, so that the terms or terms of this specification are intended to be interpreted by a skilled person based on the teachings and guidance.

本發明之寬度及範疇不應由任何上述示例性態樣限制，但應僅根據隨後申請專利範圍及其等效物來定義。The breadth and scope of the present invention should not be limited by any of the above exemplary aspects, but should be defined only in accordance with the scope of the subsequent claims and their equivalents.

本申請案全文中引用的所有引用參考文獻(包括文獻參考、美國或外國專利或專利申請案、及網站)之內容以引用之方式明確地併入本文中，如同其全文出於任何目的而在本文書寫般，其中引用的參考文獻亦如此。在出現任何不一致之情況下，以本文中按字面揭示的材料為準。The contents of all cited references (including literature references, U.S. or foreign patents or patent applications, and websites) cited throughout this application are expressly incorporated herein by reference as if written in their entirety for any purpose, including the references cited therein. In the event of any inconsistency, the material literally disclosed herein shall prevail.

圖1係來自特定腫瘤樣本之CD4 T細胞、CD8 T細胞、樹突細胞(DC)、巨噬細胞、單核細胞、初始(naïve) B-細胞、NK細胞及調節T細胞(Tregs)中之殺手細胞凝集素樣受體B1蛋白(KLRB1/CD161)、PDCD1、LAG3、CTLA4、HAVCR2及TIGIT之scRNA-seq基因表現概況之小提琴圖，如所示。Figure 1 is a violin plot of scRNA-seq gene expression profiles of killer cell lectin-like receptor B1 protein (KLRB1/CD161), PDCD1, LAG3, CTLA4, HAVCR2, and TIGIT in CD4 T cells, CD8 T cells, dendritic cells (DCs), macrophages, monocytes, naïve B-cells, NK cells, and regulatory T cells (Tregs) from specific tumor samples, as shown.

圖2係特定腫瘤樣本之初始樣CD8 T細胞、早期活性CD8 T細胞、效應子記憶CD8 T細胞、CD8 Tpex細胞及CD8 Tex細胞中之CD161、PDCD1、LAG3、CTLA4、HAVCR2、及TIGIT之scRNA-seq基因表現之小提琴圖，如所示。FIG2 is a violin plot of scRNA-seq gene expression of CD161, PDCD1, LAG3, CTLA4, HAVCR2, and TIGIT in naive-like CD8 T cells, early activated CD8 T cells, effector memory CD8 T cells, CD8 Tpex cells, and CD8 Tex cells of specific tumor samples, as shown.

圖3係對在CHOK1細胞上表現之人類及食蟹獼猴CD161蛋白具有交叉反應性之抗CD161陽性命中/抗體之基於細胞之結合之圖形表示，藉由流動式細胞測量術測定。FIG. 3 is a graphical representation of cell-based binding of anti-CD161 positive hits/antibodies cross-reactive to human and cynomolgus macaque CD161 proteins expressed on CHOK1 cells, as determined by flow cytometry.

圖4係ELISA分析中特異性抗CD161抗體對人類CD161 (hCD161)蛋白之結合之圖形表示。FIG. 4 is a graphical representation of the binding of specific anti-CD161 antibodies to human CD161 (hCD161) protein in ELISA analysis.

圖5A至圖5C係利用同型物對照(圖5A)或抗CD161陽性抗體(01D17，圖5B；及03K18，圖5C)之基於細胞之結合之FACS圖。圖5D係各種抗CD161抗體對表現人類CD161之CHOK1細胞之結合之圖形表示。Figures 5A to 5C are FACS plots of cell-based binding using isotype controls (Figure 5A) or anti-CD161 positive antibodies (01D17, Figure 5B; and 03K18, Figure 5C). Figure 5D is a graphical representation of the binding of various anti-CD161 antibodies to CHOK1 cells expressing human CD161.

圖6係人類CLEC2D多聚物與表現人類CD161之CHOK1細胞之結合之圖形表示。HP-3G10 (抗CD161參考單株抗體)，其阻斷hCLEC2D多聚物與表現hCD161之CHOK1細胞之結合。Figure 6 is a graphical representation of the binding of human CLEC2D multimers to CHOK1 cells expressing human CD161. HP-3G10 (anti-CD161 reference monoclonal antibody) blocks the binding of hCLEC2D multimers to CHOK1 cells expressing hCD161.

圖7係說明使用基於細胞之分析之抗CD161抗體之hCD161:hCLEC2D相互作用之阻斷百分比之條形圖。FIG. 7 is a bar graph illustrating the percent blocking of hCD161:hCLEC2D interaction using anti-CD161 antibodies in a cell-based assay.

圖8係顯示藉由各種抗CD161抗體濃度依賴性抑制hCD161:hCLE2CD之相互作用之ELISA分析之圖形表示。Figure 8 is a graphical representation of an ELISA analysis showing concentration-dependent inhibition of hCD161:hCLE2CD interaction by various anti-CD161 antibodies.

圖9係繪示在與各種抗CD161抗體接觸後SEB (金黃色葡萄球菌腸毒素B)刺激之健康人類供體PBMC中之IL-2細胞激素釋放之條形圖，藉由ELISA測定。FIG. 9 is a bar graph showing IL-2 cytokine release in SEB (Staphylococcus aureus enterotoxin B) stimulated healthy human donor PBMCs following exposure to various anti-CD161 antibodies, as measured by ELISA.

圖10係說明在用各種抗CD161抗體處理後藉由與Raji細胞之T細胞接合劑活化之表現hCD161之Jurkat細胞中之IL-2細胞激素釋放之條形圖，藉由ELISA測定。Figure 10 is a bar graph illustrating IL-2 cytokine release in hCD161 expressing Jurkat cells activated by T cell binder with Raji cells after treatment with various anti-CD161 antibodies, as measured by ELISA.

圖11係受CD161-CLEC2D相互作用影響的TCR活化之示意圖，其使用Jurkat-MART-1 TCR細胞及MeWo細胞作為實例。Figure 11 is a schematic diagram of TCR activation affected by CD161-CLEC2D interaction, using Jurkat-MART-1 TCR cells and MeWo cells as examples.

圖12係在不同濃度之本文描述之抗CD161抗體存在下與過度表現hCLEC2D-GFP之MeWo細胞(HLA；A*0201)接觸的MART1 TCR特異性T細胞中之TCR活化(藉由IL-2釋放測定，pg/mL)之圖形表示。頂部虛線(1)表示使Jurkat-MART1 TCR CD161細胞與模擬(GFP) MeWo細胞(對照，對TCR活性無抑制)接觸時之IL-2釋放程度。下部虛線(2)表示使Jurkat-MART1 TCR CD161細胞在不存在抗CD161抗體(對照，TCR活性之抑制)下與過度表現hCLEC2D-GFP之MeWo細胞接觸時之IL2釋放程度。Figure 12 is a graphical representation of TCR activation (as measured by IL-2 release, pg/mL) in MART1 TCR-specific T cells contacted with MeWo cells overexpressing hCLEC2D-GFP (HLA; A*0201) in the presence of different concentrations of anti-CD161 antibodies described herein. The top dashed line (1) represents the level of IL-2 release when Jurkat-MART1 TCR CD161 cells are contacted with mock (GFP) MeWo cells (control, no inhibition of TCR activity). The lower dashed line (2) shows the level of IL2 release when Jurkat-MART1 TCR CD161 cells were brought into contact with MeWo cells overexpressing hCLEC2D-GFP in the absence of anti-CD161 antibody (control, inhibition of TCR activity).

圖13係對TALL-104效應細胞(其表現CD161)之劑量依賴性抗CD161抗體結合之圖形表示。Figure 13 is a graphical representation of the dose-dependent anti-CD161 antibody binding to TALL-104 effector cells expressing CD161.

圖14A至圖14C提供細胞毒性分析之示意圖(圖14A)及顯示該細胞毒性分析能夠偵測MeWo細胞(圖14B)及PC3細胞(圖14C)之TALL-104細胞溶解之變化之初步數據。Figures 14A to 14C provide a schematic diagram of the cytotoxicity assay (Figure 14A) and preliminary data showing that the cytotoxicity assay is capable of detecting changes in TALL-104 cytolysis in MeWo cells (Figure 14B) and PC3 cells (Figure 14C).

圖15係過度表現人類CLEC2D之PC3靶細胞在漸增劑量之抗CD161單株抗體之存在下之TALL-104效應細胞細胞溶解(%)之圖形表示。頂部虛線(1)表示模擬-PC3細胞(其不表現人類CLEC2D，且因此不抑制TALL-104效應細胞)之TALL-104細胞溶解程度(%)。下部虛線(2)表示過度表現CLEC2D之PC3靶細胞在不存在抗CD161抗體下之TALL-104細胞溶解程度(%)。Figure 15 is a graphical representation of TALL-104 effector cell lysis (%) of PC3 target cells overexpressing human CLEC2D in the presence of increasing doses of anti-CD161 monoclonal antibodies. The top dashed line (1) represents the degree of TALL-104 cell lysis (%) of mock-PC3 cells (which do not express human CLEC2D and therefore do not inhibit TALL-104 effector cells). The lower dashed line (2) represents the degree of TALL-104 cell lysis (%) of PC3 target cells overexpressing CLEC2D in the absence of anti-CD161 antibodies.

圖16A至圖16B係顯示初級NK細胞與PC-3 CLEC2D-OE細胞(過度表現CLEC2D；圖16A)或Raji細胞(圖16B)在抗CD161 mAb之存在下之共培養物之相對腫瘤殺死能力之點圖。*p＜0.05，****p＜0.00001，藉由單向ANOVA與多重比較測定。Figures 16A-16B are dot plots showing the relative tumoricidal capacity of co-cultures of primary NK cells and PC-3 CLEC2D-OE cells (overexpressing CLEC2D; Figure 16A) or Raji cells (Figure 16B) in the presence of anti-CD161 mAb. *p<0.05, ****p<0.00001, determined by one-way ANOVA with multiple comparisons.

圖17係在兩次瘤內注射NK細胞且投與媒劑或該抗CD161抗體12G06後NSG小鼠中之PC3-CLEC2D-OE腫瘤體積(mm³)之圖形表示。n=4/組。*p ≤ 0.05，藉由雙向ANOVA測定。FIG. 17 is a graphical representation of PC3-CLEC2D-OE tumor volume (mm³ ) in NSG mice after two intratumoral injections of NK cells and administration of vehicle or the anti-CD161 antibody 12G06. n=4/group. *p ≤ 0.05, determined by two-way ANOVA.

圖18係在共植入人類PBMC且投與媒劑或抗CD161抗體12G06後NSG小鼠中之U87腫瘤體積(mm³)之圖形表示。n=4/組。*p ≤ 0.05，藉由雙向ANOVA測定。Figure 18 is a graphical representation of U87 tumor volume (^mm3 ) in NSG mice after co-implantation with human PBMCs and administration of vehicle or anti-CD161 antibody 12G06. n=4/group. *p ≤ 0.05, determined by two-way ANOVA.

TW202509069A_113116726_SEQL.xmlTW202509069A_113116726_SEQL.xml

Claims (40)

Translated fromChinese

一種特異性結合CD161之抗體或其抗原結合部分，其包含重鏈可變區(VH)及輕鏈可變區(VL)； 其中該VH包含VH互補決定區1 (VH-CDR1)、VH-CDR2及VH-CDR3； 其中該VL包含VL-CDR1、VL-CDR2及VL-CDR3；且 其中該VH-CDR3包含選自SEQ ID NO: 3、13、23、33、43、53、63、73、83、93、103、113、123、133、143、153、163、173、183、193、203、213、223、233、243、253、263、273、283、293、303、313、323及333所示序列之胺基酸序列。An antibody or an antigen-binding portion thereof that specifically binds to CD161, comprising a heavy chain variable region (VH) and a light chain variable region (VL); wherein the VH comprises a VH complementary determining region 1 (VH-CDR1), VH-CDR2 and VH-CDR3; wherein the VL comprises VL-CDR1, VL-CDR2 and VL-CDR3; and wherein the VH-CDR3 comprises a sequence selected from SEQ ID NO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, 133, 143, 153, 163, 173, 183, 193, 203, 213, 223, 233, 243, 253, 263, 273, 283, 293, 303, 313, 323 and the amino acid sequence of the sequence shown in 333.如請求項1之抗體或其抗原結合部分，其中該VH-CDR2包含選自SEQ ID NO: 2、12、22、32、42、52、62、72、82、92、102、112、122、132、142、152、162、172、182、192、202、212、222、232、242、252、262、272、282、292、302、312、322及332所示序列之胺基酸序列。The antibody or antigen-binding portion thereof of claim 1, wherein the VH-CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322 and 332.如請求項1或2之抗體或其抗原結合部分，其中該VH-CDR1包含選自SEQ ID NO: 1、11、21、31、41、51、61、71、81、91、101、111、121、131、141、151、161、171、181、191、201、211、221、231、241、251、261、271、281、291、301、311、321及331所示序列之胺基酸序列。The antibody or antigen-binding portion thereof of claim 1 or 2, wherein the VH-CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321 and 331.如請求項1至3中任一項之抗體或其抗原結合部分，其中該VL-CDR3包含選自SEQ ID NO: 6、16、26、36、46、56、66、76、86、96、106、116、126、136、146、156、166、176、186、196、206、216、226、236、246、256、266、276、286、296、306、316、326及336所示序列之胺基酸序列。The antibody or antigen-binding portion thereof of any one of claims 1 to 3, wherein the VL-CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326 and 336.如請求項1至4中任一項之抗體或其抗原結合部分，其中該VL-CDR2包含選自SEQ ID NO: 5、15、25、35、45、55、65、75、85、95、105、115、125、135、145、155、165、175、185、195、205、215、225、235、245、255、265、275、285、295、305、315、325及335所示序列之胺基酸序列。The antibody or antigen-binding portion thereof of any one of claims 1 to 4, wherein the VL-CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325 and 335.如請求項1至5中任一項之抗體或其抗原結合部分，其中該VL-CDR1包含選自SEQ ID NO: 4、14、24、34、44、54、64、74、84、94、104、114、124、134、144、154、164、174、184、194、204、214、224、234、244、254、264、274、284、294、304、314、324及334所示序列之胺基酸序列。The antibody or antigen-binding portion thereof of any one of claims 1 to 5, wherein the VL-CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324 and 334.如請求項1至6中任一項之抗體或其抗原結合部分，其包含： (i)    包含SEQ ID NO: 1所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 2所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 3所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 4所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 5所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 6所示胺基酸序列之VL-CDR3； (ii)   包含SEQ ID NO: 91所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 92所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 93所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 94所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 95所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 96所示胺基酸序列之VL-CDR3； (iii)  包含SEQ ID NO: 111所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 112所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 113所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 114所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 115所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 116所示胺基酸序列之VL-CDR3； (iv)   包含SEQ ID NO: 141所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 142所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 143所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 144所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 145所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 146所示胺基酸序列之VL-CDR3； (v)    包含SEQ ID NO: 211所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 212所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 213所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 214所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 215所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 216所示胺基酸序列之VL-CDR3； (vi)   包含SEQ ID NO: 221所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 222所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 223所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 224所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 225所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 226所示胺基酸序列之VL-CDR3； (vii)  包含SEQ ID NO: 271所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 272所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 273所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 274所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 275所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 276所示胺基酸序列之VL-CDR3； (viii) 包含SEQ ID NO: 281所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 282所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 283所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 284所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 285所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 286所示胺基酸序列之VL-CDR3； (ix)   包含SEQ ID NO: 291所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 292所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 293所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 294所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 295所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 296所示胺基酸序列之VL-CDR3； (x)    包含SEQ ID NO: 301所示胺基酸序列之VH-CDR1、包含SEQ ID NO: 302所示胺基酸序列之VH-CDR2、包含SEQ ID NO: 303所示胺基酸序列之VH-CDR3、包含SEQ ID NO: 304所示胺基酸序列之VL-CDR1、包含SEQ ID NO: 305所示胺基酸序列之VL-CDR2、及包含SEQ ID NO: 306所示胺基酸序列之VL-CDR3；或 (xi)   (i)至(x)之任何組合。An antibody or antigen-binding portion thereof as claimed in any one of claims 1 to 6, comprising:(i)    a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 6;(ii)   a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 91, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 92, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 93, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 94, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 95, and a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 96;(iii)  A VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 111, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 112, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 113, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 114, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 116;(iv)   A VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 144, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 145, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 146;(v)    a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 213, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 216;(vi)   a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 211, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 212, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 213, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 214, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 215, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 216; 221, a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 222, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 223, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 224, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 225, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 226; (vii)  a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 271, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 272, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 273, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 274, and a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 276. 275, and a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 276;(viii) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 281, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 282, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 283, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 284, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 285, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 286;(ix)   a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 291, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 292, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 293, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 294, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 295, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 296;(x)    a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 302, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 303, a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 306; or(xi)   any combination of (i) to (x).如請求項1至7中任一項之抗體或其抗原結合部分，其中該VH包含與選自SEQ ID NO: 7、17、27、37、47、57、67、77、87、97、107、117、127、137、147、157、167、177、187、197、207、217、227、237、247、257、267、277、287、297、707、317、327及337之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列。The antibody or antigen-binding portion thereof of any one of claims 1 to 7, wherein the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337.如請求項1至8中任一項之抗體或其抗原結合部分，其中該VL包含與選自SEQ ID NO: 8、18、28、38、48、58、68、78、88、98、108、118、128、138、148、158、168、178、188、198、208、218、228、238、248、258、268、278、288、298、308、318、328及338之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列。The antibody or antigen-binding portion thereof of any one of claims 1 to 8, wherein the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328, and 338.一種特異性結合CD161之抗體或其抗原結合部分，其包含可變重(VH)域及可變輕(VL)域，其中該VH包含與選自SEQ ID NO: 7、17、27、37、47、57、67、77、87、97、107、117、127、137、147、157、167、177、187、197、207、217、227、237、247、257、267、277、287、297、707、317、327及337之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且其中該VL包含與選自SEQ ID NO: 8、18、28、38、48、58、68、78、88、98、108、118、128、138、148、158、168、178、188、198、208、218、228、238、248、258、268、278、288、298、308、318、328及338之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列。An antibody or antigen-binding portion thereof that specifically binds CD161, comprising a variable heavy (VH) domain and a variable light (VL) domain, wherein the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327, and 337; and wherein the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: The amino acid sequences of NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328 and 338 have an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity.如請求項1至10中任一項之抗體或其抗原結合部分，其中該VH包含選自SEQ ID NO: 7、17、27、37、47、57、67、77、87、97、107、117、127、137、147、157、167、177、187、197、207、217、227、237、247、257、267、277、287、297、707、317、327及337之胺基酸序列。The antibody or antigen-binding portion thereof of any one of claims 1 to 10, wherein the VH comprises an amino acid sequence selected from SEQ ID NO: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 707, 317, 327 and 337.如請求項1至11中任一項之抗體或其抗原結合部分，其中該VL包含選自SEQ ID NO: 8、18、28、38、48、58、68、78、88、98、108、118、128、138、148、158、168、178、188、198、208、218、228、238、248、258、268、278、288、298、308、318、328及338之胺基酸序列。The antibody or antigen-binding portion thereof of any one of claims 1 to 11, wherein the VL comprises an amino acid sequence selected from SEQ ID NO: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, 118, 128, 138, 148, 158, 168, 178, 188, 198, 208, 218, 228, 238, 248, 258, 268, 278, 288, 298, 308, 318, 328 and 338.如請求項1至12中任一項之抗體或其抗原結合部分，其中 (i)    該VH包含與選自SEQ ID NO: 7之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 8之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (ii)   該VH包含與選自SEQ ID NO: 97之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 98之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (iii)  該VH包含與選自SEQ ID NO: 117之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 118之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (iv)   該VH包含與選自SEQ ID NO: 147之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 148之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (v)    該VH包含與選自SEQ ID NO: 217之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 218之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (vi)   該VH包含與選自SEQ ID NO: 227之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 228之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (vii)  該VH包含與選自SEQ ID NO: 277之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 278之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (viii) 該VH包含與選自SEQ ID NO: 287之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 288之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (ix)   該VH包含與選自SEQ ID NO: 297之胺基酸序列具有80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 298之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列； (x)    該VH包含與選自SEQ ID NO: 307之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；且該VL包含與選自SEQ ID NO: 308之胺基酸序列具有至少約80%、至少約85%、至少約90%、至少約95%、至少約96%、至少約97%、至少約98%或至少約99%序列一致性之胺基酸序列；或 (xi)   (i)至(x)之任何組合。The antibody or antigen-binding portion thereof of any one of claims 1 to 12, wherein (i)   the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 7; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 8; (ii)   the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 97 has an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity; and the VL comprises an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 98; (iii)  the VH comprises an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 117; and the VL comprises an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 118 has an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity; (iv)   the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 147; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 148; (v)   the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 217 has an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity; and the VL comprises an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 218; (vi)   the VH comprises an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 227; and the VL comprises an amino acid sequence with at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NO: 228 has an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity;(vii) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 277; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 278;(viii) the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 287 has an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 288; (ix)   the VH comprises an amino acid sequence having 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 297; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 298 has at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity;(x)    the VH comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 307; and the VL comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 308; or(xi)   any combination of (i) to (x).如請求項1至13中任一項之抗體或其抗原結合部分，其包含： (i)    包含SEQ ID NO: 7所示胺基酸序列之VH、及包含SEQ ID NO: 8所示胺基酸序列之VL； (ii)   包含SEQ ID NO: 97所示胺基酸序列之VH、及包含SEQ ID NO: 98所示胺基酸序列之VL； (iii)  包含SEQ ID NO: 117所示胺基酸序列之VH、及包含SEQ ID NO: 118所示胺基酸序列之VL； (iv)   包含SEQ ID NO: 147所示胺基酸序列之VH、及包含SEQ ID NO: 148所示胺基酸序列之VL； (v)    包含SEQ ID NO: 217所示胺基酸序列之VH、及包含SEQ ID NO: 218所示胺基酸序列之VL； (vi)   包含SEQ ID NO: 227所示胺基酸序列之VH、及包含SEQ ID NO: 228所示胺基酸序列之VL； (vii)  包含SEQ ID NO: 277所示胺基酸序列之VH、及包含SEQ ID NO: 278所示胺基酸序列之VL； (viii)包含SEQ ID NO: 287所示胺基酸序列之VH、及包含SEQ ID NO: 288所示胺基酸序列之VL； (ix)   包含SEQ ID NO: 297所示胺基酸序列之VH、及包含SEQ ID NO: 298所示胺基酸序列之VL； (x)    包含SEQ ID NO: 307所示胺基酸序列之VH、及包含SEQ ID NO: 308所示胺基酸序列之VL； (xi)   (i)至(x)之任何組合。An antibody or antigen-binding portion thereof according to any one of claims 1 to 13, comprising: (i)    a VH comprising the amino acid sequence of SEQ ID NO: 7, and a VL comprising the amino acid sequence of SEQ ID NO: 8; (ii)   a VH comprising the amino acid sequence of SEQ ID NO: 97, and a VL comprising the amino acid sequence of SEQ ID NO: 98; (iii)   a VH comprising the amino acid sequence of SEQ ID NO: 117, and a VL comprising the amino acid sequence of SEQ ID NO: 118; (iv)   a VH comprising the amino acid sequence of SEQ ID NO: 147, and a VL comprising the amino acid sequence of SEQ ID NO: 148; (v)   a VH comprising the amino acid sequence of SEQ ID NO: 217, and a VL comprising the amino acid sequence of SEQ ID NO: 218; (vi)   a VH comprising the amino acid sequence of SEQ ID NO: 227, and VL comprising the amino acid sequence of SEQ ID NO: 228;(vii)  A VH comprising the amino acid sequence of SEQ ID NO: 277, and VL comprising the amino acid sequence of SEQ ID NO: 278;(viii)  A VH comprising the amino acid sequence of SEQ ID NO: 287, and VL comprising the amino acid sequence of SEQ ID NO: 288;(ix)   A VH comprising the amino acid sequence of SEQ ID NO: 297, and VL comprising the amino acid sequence of SEQ ID NO: 298;(x)   A VH comprising the amino acid sequence of SEQ ID NO: 307, and VL comprising the amino acid sequence of SEQ ID NO: 308;(xi)   Any combination of (i) to (x).一種抗體或其抗原結合部分，其結合與如請求項1至14中任一項之抗體或其抗原結合部分相同的抗原決定基。An antibody or antigen-binding portion thereof, which binds to the same antigenic determinant as the antibody or antigen-binding portion thereof of any one of claims 1 to 14.一種抗體或其抗原結合部分，其與如請求項1至14中任一項之抗體或其抗原結合部分交叉競爭結合CD161。An antibody or an antigen-binding portion thereof, which cross-competitively binds to CD161 with the antibody or antigen-binding portion thereof of any one of claims 1 to 14.如請求項1至16中任一項之抗體或其抗原結合部分，其以小於約1000 nM、小於約500 nM、小於約100 nM、小於約50 nM或小於約10 nM之K_D結合CD161。The antibody, or antigen-binding portion thereof, of any one of claims 1 to 16, which binds CD161 with a_KD of less than about 1000 nM, less than about 500 nM, less than about 100 nM, less than about 50 nM, or less than about 10 nM.如請求項1至17中任一項之抗體或其抗原結合部分，其以小於約50 nM之K_D結合CD161。The antibody or antigen-binding portion thereof of any one of claims 1 to 17, which binds CD161 with a_KD of less than about 50 nM.如請求項1至18中任一項之抗體或其抗原結合部分，其以小於約10 nM之K_D結合CD161。The antibody or antigen-binding portion thereof of any one of claims 1 to 18, which binds CD161 with a_KD of less than about 10 nM.如請求項1至19中任一項之抗體或其抗原結合部分，其抑制CD161與C型凝集素域家族2成員D (CLEC2D)之間之相互作用。The antibody or antigen-binding portion thereof of any one of claims 1 to 19, which inhibits the interaction between CD161 and C-type lectin domain family 2 member D (CLEC2D).如請求項1至20中任一項之抗體或其抗原結合部分，其能夠誘導或增強免疫細胞產生一或多種細胞激素。The antibody or antigen-binding portion thereof of any one of claims 1 to 20, which is capable of inducing or enhancing immune cells to produce one or more cytokines.如請求項21之抗體或其抗原結合部分，其中該一或多種細胞激素包含IL2、TNFa、IFNg或其任何組合。The antibody or antigen-binding portion thereof of claim 21, wherein the one or more cytokines comprise IL2, TNFa, IFNg or any combination thereof.如請求項1至22中任一項之抗體或其抗原結合部分，其中該抗體之該抗原結合部分包含VHH、vNAR、微體(microbody)、奈米抗體(nanobody)、scFv或其任何組合。The antibody or antigen-binding portion thereof of any one of claims 1 to 22, wherein the antigen-binding portion of the antibody comprises VHH, vNAR, microbody, nanobody, scFv or any combination thereof.一種多特異性抗體，其包含如請求項1至23中任一項之抗體或其抗原結合部分。A multispecific antibody comprising the antibody of any one of claims 1 to 23 or an antigen-binding portion thereof.一種雙特異性抗體，其包含如請求項1至23中任一項之抗體或其抗原結合部分。A bispecific antibody comprising the antibody of any one of claims 1 to 23 or an antigen-binding portion thereof.一種核酸分子或一組核酸分子，其編碼如請求項1至23中任一項之抗體或其抗原結合部分、如請求項24之多特異性抗體或如請求項25之雙特異性抗體。A nucleic acid molecule or a group of nucleic acid molecules encoding the antibody or antigen-binding portion thereof of any one of claims 1 to 23, the multispecific antibody of claim 24, or the bispecific antibody of claim 25.一種載體或一組載體，其包含如請求項26之核酸分子或一組核酸分子。A vector or a group of vectors comprising the nucleic acid molecule or a group of nucleic acid molecules as claimed in claim 26.如請求項36之載體或一組載體，其係病毒載體。The vector or a group of vectors as claimed in claim 36 is a viral vector.一種宿主細胞，其包含如請求項26之核酸分子或一組核酸分子或如請求項27或28之載體或一組載體。A host cell comprising the nucleic acid molecule or a group of nucleic acid molecules of claim 26 or the vector or a group of vectors of claim 27 or 28.一種醫藥組合物，其包含如請求項1至23中任一項之抗體或其抗原結合部分、如請求項24之多特異性抗體、如請求項25之雙特異性抗體、如請求項26之核酸分子或一組核酸分子、如請求項27或28之載體或一組載體、或如請求項29之宿主細胞及醫藥上可接受之載劑。A pharmaceutical composition comprising an antibody or an antigen-binding portion thereof as described in any one of claims 1 to 23, a multispecific antibody as described in claim 24, a bispecific antibody as described in claim 25, a nucleic acid molecule or a group of nucleic acid molecules as described in claim 26, a vector or a group of vectors as described in claim 27 or 28, or a host cell as described in claim 29, and a pharmaceutically acceptable carrier.一種治療有需要個體之疾病或病症之方法，該方法包括對該個體投與如請求項1至23中任一項之抗體或其抗原結合部分、如請求項24之多特異性抗體、如請求項25之雙特異性抗體、如請求項26之核酸分子或一組核酸分子、如請求項27或28之載體或一組載體、如請求項29之宿主細胞或如請求項30之醫藥組合物。A method for treating a disease or condition in an individual in need thereof, the method comprising administering to the individual an antibody or antigen-binding portion thereof as described in any one of claims 1 to 23, a multispecific antibody as described in claim 24, a bispecific antibody as described in claim 25, a nucleic acid molecule or a group of nucleic acid molecules as described in claim 26, a vector or a group of vectors as described in claim 27 or 28, a host cell as described in claim 29, or a pharmaceutical composition as described in claim 30.如請求項31之方法，其中該疾病或病症包含癌症。The method of claim 31, wherein the disease or condition comprises cancer.一種誘導有需要個體中免疫反應之方法，該方法包括對該個體投與如請求項1至23中任一項之抗體或其抗原結合部分、如請求項24之多特異性抗體、如請求項25之雙特異性抗體、如請求項26之核酸分子或一組核酸分子、如請求項27或28之載體或一組載體、如請求項29之宿主細胞或如請求項30之醫藥組合物。A method for inducing an immune response in an individual in need thereof, the method comprising administering to the individual an antibody or antigen-binding portion thereof as described in any one of claims 1 to 23, a multispecific antibody as described in claim 24, a bispecific antibody as described in claim 25, a nucleic acid molecule or a group of nucleic acid molecules as described in claim 26, a vector or a group of vectors as described in claim 27 or 28, a host cell as described in claim 29, or a pharmaceutical composition as described in claim 30.如請求項33之方法，其中該個體罹患癌症。The method of claim 33, wherein the individual suffers from cancer.一種治療有需要個體之癌症之方法，該方法包括對該個體投與如請求項1至23中任一項之抗體或其抗原結合部分、如請求項24之多特異性抗體、如請求項25之雙特異性抗體、如請求項26之核酸分子或一組核酸分子、如請求項27或28之載體或一組載體、如請求項29之宿主細胞或如請求項30之醫藥組合物。A method for treating cancer in an individual in need thereof, the method comprising administering to the individual an antibody or antigen-binding portion thereof as described in any one of claims 1 to 23, a multispecific antibody as described in claim 24, a bispecific antibody as described in claim 25, a nucleic acid molecule or a group of nucleic acid molecules as described in claim 26, a vector or a group of vectors as described in claim 27 or 28, a host cell as described in claim 29, or a pharmaceutical composition as described in claim 30.如請求項32、34及35中任一項之方法，其中該癌症選自聽神經瘤、急性淋巴球性白血病、急性淋巴球性白血病、腺癌、及泌尿系統癌、及上皮細胞癌、血管肉瘤、星形細胞瘤、基底細胞癌、膽管癌、膽道癌、膀胱癌、骨癌、腦癌、腦幹神經膠質瘤、乳癌、支氣管癌、伯奇氏淋巴瘤(Burkitt's lymphoma)及邊緣區B細胞淋巴瘤、腎上腺癌、肛門區癌、消化系統癌、內分泌系統癌、食道癌、副甲狀腺癌、陰莖癌、呼吸系统癌、小腸癌、輸尿管癌、尿道癌、子宮頸癌、子宮內膜癌、輸卵管癌、腎盂癌、陰道癌、外陰癌、中樞神經系統(CNS)癌症、子宮頸癌、軟骨肉瘤、脊索瘤、絨毛膜癌、慢性白血病、慢性淋巴球性白血病、慢性髓細胞性(粒細胞性)白血病、結腸癌、結腸肉瘤、結腸直腸癌、結締組織癌、顱咽管瘤、囊腺癌、胚胎癌、子宮內膜癌、內皮肉瘤、環境誘導型癌症(包括彼等由石棉誘導者)、室管膜瘤、表皮樣癌、上皮樣癌、食道癌、食道上皮細胞癌、尤因氏肉瘤(Ewing's tumor)、眼癌、纖維肉瘤、胃癌(gastric cancer)、胃腸癌、生殖細胞腫瘤、神經膠質母細胞瘤(例如多形性神經膠質母細胞瘤)、神經膠質瘤、頭頸癌、重鏈病(heavy chain disease)、血管母細胞瘤、肝癌瘤、霍奇金氏病(Hodgkin's disease)、上皮內贅瘤、卡波西氏肉瘤(Kaposi's sarcoma)、腎癌(例如腎細胞癌(RCC))、喉頭癌、平滑肌肉瘤、白血病、脂肪肉瘤、肝癌、肺癌(lung cancer) (小細胞、大細胞)、肺癌(lung carcinoma)、淋巴管內皮肉瘤、淋巴管肉瘤、被套細胞淋巴瘤、髓質癌、神經管胚細胞瘤、黑色素瘤、腦膜瘤(menangioma)、間皮瘤、多發性骨髓瘤、骨髓母細胞前髓細胞骨髓單核球性單核細胞紅血球性白血病、黏液肉瘤、鼻咽癌、中樞神經系統(CNS)贅瘤、神經母細胞瘤、非霍奇金氏病(non-Hodgkin's disease)、非小細胞肺癌(non-small cell lung cancer；NSCLC)、非小細胞肺上皮細胞癌、寡樹突神經膠質瘤、口腔癌(例如唇、舌、口及咽)、成骨肉瘤、骨肉瘤、卵巢癌、胰臟癌、乳突腺癌、乳突癌、小兒肉瘤、松果腺瘤(pinealoma)、垂體腺瘤、真性紅血球增多症(Polycythemia vera)、淋巴瘤、原發性CNS淋巴瘤、前列腺癌(例如激素難治性前列腺腺癌)、直腸癌、腎癌(例如透明細胞癌)、視網膜母細胞瘤、橫紋肌肉瘤、肉瘤、軟組織肉瘤、皮脂腺癌、精細胞瘤、鼻腔鼻竇自然殺手(sinonasal natural killer)、皮膚癌、小細胞肺癌(SCLC)、兒童實體腫瘤、脊髓軸瘤、鱗狀細胞癌、鱗狀細胞上皮細胞癌、胃癌(stomach cancer)、汗腺癌、滑膜瘤、睾丸癌、甲狀腺癌、腫瘤血管生成、子宫癌、病毒相關癌症或病毒起源之癌症(例如，人類乳頭瘤病毒(HPV相關或來源腫瘤))、華氏巨球蛋白血症(Waldenstrom's macroglobulinemia)及威爾姆氏腫瘤(Wilm's tumor)；及該等癌症之任何組合。The method of any one of claims 32, 34 and 35, wherein the cancer is selected from acoustic neuroma, acute lymphocytic leukemia, acute lymphocytic leukemia, adenocarcinoma, and urinary system cancer, and epithelial cell carcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bile duct cancer, bladder cancer, bone cancer, brain cancer, brain stem neurofibroma, breast cancer, bronchial carcinoma, Birch's lymphoma (Burkitt's lymphoma) and marginal B-cell lymphoma, adrenal cancer, anal cancer, digestive system cancer, endocrine system cancer, esophageal cancer, parathyroid cancer, penile cancer, respiratory system cancer, small intestine cancer, ureteral cancer, urethral cancer, cervical cancer, endometrial cancer, fallopian tube cancer, renal pelvis cancer, vaginal cancer, vulvar cancer, central nervous system (CNS) cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, Chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, colon cancer, colon sarcoma, colorectal cancer, connective tissue cancer, cranio-pharyngioma, cystadenocarcinoma, embryonal carcinoma, endometrial carcinoma, endothelial sarcoma, environmentally induced cancers (including those induced by asbestos), ependymoma, epidermoid carcinoma, epithelioid carcinoma, esophageal cancer, esophageal epithelial cell carcinoma, Ewing's sarcoma tumor), eye cancer, fibrosarcoma, gastric cancer, gastrointestinal cancer, germ cell tumor, neuroglioblastoma (e.g., multiforme neuroglioblastoma), neuroglioma, head and neck cancer, heavy chain disease, hemangioblastoma, hepatocarcinoma, Hodgkin's disease, intraepithelial neoplasia, Kaposi's sarcoma, kidney cancer (e.g., renal cell carcinoma (RCC)), laryngeal cancer, leiomyosarcoma, leukemia, liposarcoma, liver cancer, lung cancer (small cell, large cell), lung cancer (lung carcinoma), lymphangioendothelioma, lymphangiosarcoma, mantle cell lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloblastic promyelocytic myelomonocytic monocytic erythrocytic leukemia, myxosarcoma, nasopharyngeal carcinoma, central nervous system (CNS) neoplasms, neuroblastoma, non-Hodgkin's disease, non-small cell lung cancer cancer; NSCLC), non-small cell lung epithelial carcinoma, oligodendroglioma, oral cancer (e.g., lip, tongue, mouth, and pharynx), osteogenic sarcoma, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pediatric sarcoma, pinealoma, pituitary adenoma, polycythemia vera, lymphoma, primary CNS lymphoma, prostate cancer (e.g., hormone-refractory prostate adenocarcinoma), rectal cancer, kidney cancer (e.g., clear cell carcinoma), retinoblastoma, rhabdomyosarcoma, sarcoma, soft tissue sarcoma, sebaceous gland carcinoma, spermatoma, sinonasal natural killer (sinonasal natural killer), killer), skin cancer, small cell lung cancer (SCLC), solid tumors of children, spinal cord tumors, squamous cell carcinoma, squamous cell epithelial carcinoma, stomach cancer, sweat gland carcinoma, synovial tumor, testicular cancer, thyroid cancer, tumor angiogenesis, uterine cancer, virus-related cancer or cancer of viral origin (e.g., human papillomavirus (HPV-related or derived tumors)), Waldenstrom's macroglobulinemia, and Wilm's tumor; and any combination of these cancers.一種治療有需要個體之傳染病之方法，該方法包括對該個體投與如請求項1至23中任一項之抗體或其抗原結合部分、如請求項24之多特異性抗體、如請求項25之雙特異性抗體、如請求項26之核酸分子或一組核酸分子、如請求項27或28之載體或一組載體、如請求項29之宿主細胞或如請求項30之醫藥組合物。A method for treating an infectious disease in an individual in need thereof, the method comprising administering to the individual an antibody or an antigen-binding portion thereof as described in any one of claims 1 to 23, a multispecific antibody as described in claim 24, a bispecific antibody as described in claim 25, a nucleic acid molecule or a group of nucleic acid molecules as described in claim 26, a vector or a group of vectors as described in claim 27 or 28, a host cell as described in claim 29, or a pharmaceutical composition as described in claim 30.如請求項37之方法，其中該傳染病包含： (i)    流行性感冒、皰疹、梨形鞭毛蟲屬(Giardia)、瘧疾、利什曼原蟲(Leishmania)或其任何組合之感染； (ii)   人類免疫缺陷病毒(HIV)、肝炎病毒、皰疹病毒、腺病毒、流行性感冒病毒、黃病毒科、伊科病毒(echovirus)、鼻病毒、柯薩奇病毒(coxsackie virus)、冠狀病毒、呼吸道融合病毒、腮腺炎病毒、輪狀病毒、麻疹病毒、蕁麻疹病毒、小病毒、牛痘病毒、HTLV病毒、登革熱病毒(dengue virus)、乳頭瘤病毒、軟疣病毒、脊髓灰質炎病毒、狂犬病病毒、JC病毒、或蟲媒病毒腦炎病毒、或其任何組合之感染； (iii)  披衣菌、立克次體細菌、分枝桿菌、葡萄球菌、鏈球菌、肺炎球菌、腦膜炎雙球菌、淋球菌、克雷伯氏菌(klebsiella)、變形桿菌屬、沙雷氏菌(serratia)、假單胞菌、軍團菌屬(legionella)、白喉桿菌(diphtheria)、沙門氏菌(salmonella)、桿菌、霍亂、破傷風、肉毒症、炭疽、瘟疫、鉤端螺旋體病、及萊姆氏病(Lyme’s disease)細菌、或其任何組合之感染； (iv)   念珠菌屬(Candida)、新型隱球菌(Cryptococcus neoformans)、麴菌屬(Aspergillus)、毛黴目(Mucorales)屬、申克氏胞絲菌(Sporothrix schenkii)、皮炎芽生菌(Blastomyces dermatitidis)、巴西副球孢子菌(Paracoccidioides brasiliensis)、粗球黴菌(Coccidioides immitis)、或莢膜組織漿菌(Histoplasma capsulatum)、或其任何組合之感染； (v)    溶組織內阿米巴(Entamoeba histolytica)、結腸小袋纖毛蟲(Balantidium coli)、福氏耐格里阿米巴原蟲(Naegleriafowleri)、棘阿米巴科(Acanthamoeba sp.)、蘭氏賈第鞭毛蟲(Giardia lambia)、隱孢子蟲(Cryptosporidium sp.)、卡式肺囊蟲(Pneumocystis carinii)、間日瘧原蟲(Plasmodium vivax)、巴貝斯原蟲(Babesia microti)、布氏錐蟲(Trypanosoma brucei)、克氏錐蟲(Trypanosoma cruzi)、杜氏利什曼原蟲(Leishmania donovani)、岡地弓漿蟲病(Toxoplasma gondi)、或巴西日圓線蟲(Nippostrongylus brasiliensis)、或其任何組合之感染；或 (vi)   (i)至(v)之任何組合。The method of claim 37, wherein the infectious disease comprises: (i) infection by influenza, herpes, Giardia, malaria, Leishmania, or any combination thereof; (ii) infection by human immunodeficiency virus (HIV), hepatitis virus, herpes virus, adenovirus, influenza virus, Flaviviridae, echovirus, rhinovirus, coxsackie virus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum virus, polio virus, rabies virus, JC virus, or arbovirus encephalitis virus, or any combination thereof; (iii) (iv) infections caused by bacteria of the genus Candida, Cryptococcusneoformans , Aspergillus, Mucorales,Schenckia , and Lyme's disease; (v) infections caused by bacteria of the genus Candida,Cryptococcusneoformans ,Aspergillus, Mucorales, and Schenckia; (vi) infections caused by bacteria of the genus Cryptococcus,Aspergillus,Mucorales , andSchenckia ; (v) infection withEntamoebahistolytica ,Balantidium coli, Naegleria fowleri, Acanthamoeba sp.,Giardialambia ,Cryptosporidiumsp. , Pneumocystiscarinii ,Plasmodium vivax,vivax ),Babesia microti ,Trypanosoma brucei ,Trypanosoma cruzi ,Leishmania donovani ,Toxoplasma gondi , orNippostrongylus brasiliensis , or any combination thereof; or (vi) any combination of (i) to (v).一種治療有需要個體之自體免疫疾病之方法，該方法包括對該個體投與如請求項1至23中任一項之抗體或其抗原結合部分、如請求項24之多特異性抗體、如請求項25之雙特異性抗體、如請求項26之核酸分子或一組核酸分子、如請求項27或28之載體或一組載體、如請求項29之宿主細胞或如請求項30之醫藥組合物。A method for treating an autoimmune disease in an individual in need thereof, the method comprising administering to the individual an antibody or antigen-binding portion thereof as described in any one of claims 1 to 23, a multispecific antibody as described in claim 24, a bispecific antibody as described in claim 25, a nucleic acid molecule or a group of nucleic acid molecules as described in claim 26, a vector or a group of vectors as described in claim 27 or 28, a host cell as described in claim 29, or a pharmaceutical composition as described in claim 30.一種活化免疫細胞之方法，該方法包括使該免疫細胞與如請求項1至23中任一項之抗體或其抗原結合部分、如請求項24之多特異性抗體、如請求項25之雙特異性抗體、如請求項26之核酸分子或一組核酸分子、如請求項27或28之載體或一組載體、如請求項29之宿主細胞或如請求項30之醫藥組合物接觸。A method for activating an immune cell, the method comprising contacting the immune cell with an antibody or an antigen-binding portion thereof as described in any one of claims 1 to 23, a multispecific antibody as described in claim 24, a bispecific antibody as described in claim 25, a nucleic acid molecule or a group of nucleic acid molecules as described in claim 26, a vector or a group of vectors as described in claim 27 or 28, a host cell as described in claim 29, or a pharmaceutical composition as described in claim 30.