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.2020 Jan;20(1):64-74.
doi: 10.1111/ajt.15501. Epub 2019 Jul 15.

Radiation and host retinoic acid signaling promote the induction of gut-homing donor T cells after allogeneic hematopoietic stem cell transplantation

Affiliations

Radiation and host retinoic acid signaling promote the induction of gut-homing donor T cells after allogeneic hematopoietic stem cell transplantation

Jianwei Zheng et al. Am J Transplant.2020 Jan.

Abstract

Intestinal graft-versus-host disease (GVHD) remains a devastating complication after allogeneic hematopoietic stem cell transplantation (HSCT). Although it has been well established that gut-tropic donor T cells expressing integrin α4β7 are required to cause intestinal damage, the factors that control the induction of this pathogenic T cell population remain to be identified. Retinoic acid (RA) plays an important role in inducing α4β7 expression on T cells. In this study, we showed that gene expression of retinaldehyde dehydrogenase, the key enzyme involved in RA biosynthesis, is significantly increased in the spleen and mesenteric lymph nodes (MLNs) of irradiated mice. In a C57BL/6-into-B6D2F1 allogeneic HSCT model, irradiation significantly increased the induction of α4β7+ -donor T cells in mesenteric lymph nodes and spleen. Furthermore, we found that the RA pathway modulates the ability of dendritic cells to imprint gut-homing specificity on alloreactive T cells. We also showed that host dendritic cell RA signaling influences GVHD risk. Our studies identified radiation and recipient RA signaling as 2 primary factors that dictate the magnitude of gut-homing donor T cell induction after allogeneic HSCT. Attenuating radiation-associated inflammation and modulating host RA signaling represent feasible strategies to mitigate intestinal GVHD by reducing gut-seeking pathogenic donor T cells.

Keywords: animal models: murine; basic (laboratory) research/science; bone marrow/hematopoietic stem cell transplantation; graft-versus-host disease (GVHD); immune regulation; immunobiology.

© 2019 The American Society of Transplantation and the American Society of Transplant Surgeons.

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Conflict of interest statement

DISCLOSURE

The authors of this manuscript have no conflicts of interest to disclose as described by theAmerican Journal of Transplantation.

Figures

Figure 1:
Figure 1:. Irradiation increases RALDH gene expression in the MLNs and spleen.
Naïve Balb/cJ mice received total body irradiation (TBI) at 900 cGy. 24 and 72 hours after irradiation, MLNs (A), spleen (B), and liver (C) were excised from these mice. RNA was extracted from these tissues and gene expression ofAldh1a1,Aldh1a2, andAldh1a3 was analyzed by real-time q-PCR. Data are normalized for β2-microglobulin RNA and presented as fold increase over gene expression in non-irradiated animals. MLNs are pooled from 3 mice in each group and data are the cumulative results from 3 independent experiments. Data for spleen are the cumulative results from 2 experiments (n = 6 per group). Data for liver are derived from one representative of two experiments (n = 3 per group). Statistics: *P ≤ .05, ****P ≤ .0001.
Figure 2:
Figure 2:. Radiation enhances RALDH enzyme activity and the induction of gut-homing donor T cells early after allogeneic BMT.
(A-D) Balb/cJ mice received TBI at 900 cGy and were euthanized on day 2 after irradiation. 0.5×106 MLN or spleen cells were suspended in ALDEFLUOR assay buffer containing the activated ALDEFLUOR substrate and incubated for 45 minutes at 37 °C. Cells were then stained with anti-CD45, anti-CD11c and anti-MHC-II antibodies and 7-AAD. Live (7-AAD–) CD45+CD11c+MHC-II+ALDEFLUOR+ cells were quantified by flow cytometry. Representative contour plots depicting ALDEFLUOR+ DCs from spleen (A) and MLN (C) are shown. (B, D) Mean (± SEM) percentage of ALDEFLUOR+ DCs from spleen (B) and MLN (D). Data are the cumulative results from 5 independent experiments;(E-H) Lethally irradiated (1300 cGy) or non-irradiated B6D2F1 mice were transplanted with 5 × 106 B6 BM together with B6 splenocytes (adjusted to yield a T-cell dose of 4 × 106). Mice in both groups were euthanized on day 4 after transplantation. The expression of integrin α4β7 on donor T cells isolated from MLNs and spleen of recipient mice was examined by gating on H-2b+H2d– CD4+ or H-2b+H2d– CD8+ donor T cells. (E, G) Representative contour plots for α4β7 expression on donor T cells isolated from MLNs (E) and spleen (G) are shown. (F, H) Mean (± SEM) percentage of α4β7+ donor T cells isolated from MLNs (F) and spleen (H) (n = 4 mice per group). Data are derived from one representative of 3 experiments. Statistics: **P ≤ .01, ***P ≤ .001, ****P ≤ .0001.
Figure 3:
Figure 3:. RA-exposed DCs show increased ability to stimulate the proliferation of alloreactive T cells with heightened gut-homing potential.
Naïve Balb/cJ mice received TBI and were intraperitoneally injected with either DMSO or RA (900 μg). After 18 hours, highly purified CD11c+ DCs were isolated from the spleens of these mice. 5 × 104 DMSO- or RA-exposed DCs were co-cultured with 1 × 105 purified B6 splenic pan-T cells labeled with CellTrace Violet dye. Four days later, cells were harvested for flow cytometric analysis.(A, B) Representative flow plots for CCR9 and α4β7 expression on CD4+ and CD8+ alloreactive T cells.(C) Mean (± SEM) percentage of alloreactive CD4+ and CD8+ T cells expressing gut-homing molecules CCR9 and α4β7 from triplicate wells.(D) Mean (± SEM) percentage of proliferating (CellTrace Violet low) CD4+ and CD8+ T cells from triplicate wells.(E) Mean (± SEM) percentage of CD4+IFN-γ+ and CD8+IFN-γ+ cells from triplicate wells as assessed by intracellular cytokine staining. Data are derived from one representative of at least three experiments. Statistics: *P ≤ .05, **P ≤ .01, ***P ≤ .001, ****P ≤ .0001.
Figure 4:
Figure 4:. Abrogating RA signaling in DCs inhibits their ability to induce gut-homing molecule expression on alloreactive T cells.
Purified CD11c+ DCs were isolated from the MLNs of vitamin A normal (VAN) or vitamin A deficient (VAD) mice. 5 × 104 of these DCs were co-cultured with 1 × 105 purified spleen pan-T cells labeled with CellTrace Violet dye from naïve B6 mice. Four days later, cells were harvested for flow cytometric analysis.(A) Representative flow plots for α4β7 expression on CD4+ and CD8+ alloreactive T cells.(B) Mean (± SEM) percentage of proliferating alloreactive CD4+α4β7+ and CD8+α4β7+ T cells from triplicate wells. Data are derived from one representative of three experiments.(C) Naïve Balb/cJ mice received TBI and were intraperitoneally injected with either DMSO or BMS493 (440 μg). After 18 hours, purified CD11c+ DCs were isolated from the spleens of these mice. 5 × 104 DMSO- or BMS493-exposed DCs were co-cultured with 1 × 105 purified spleen pan-T cells labeled with CellTrace Violet dye from naïve B6 animals. Four days later, cells were harvested for flow cytometric analysis. Mean (± SEM) percentage of proliferating alloreactive CD4+ and CD8+ T cells expressing α4β7 from 3 wells is shown. Data are derived from one representative of two experiments.(D) Naïve VAD mice received 4 doses of either DMSO or RA (450 μg/mouse) by intraperitoneal injections from day −3 to day 0 of BMT. Lethally irradiated VAN and DMSO- or RA-treated VAD mice were transplanted with B6 BM (5 × 106) and splenic T cells (adjusted to yield a T-cell dose of 1.5 × 106). Recipient mice were euthanized on day 4 after BMT. Percentage of CCR9 and α4β7 positive donor T cells (H-2b+CD4+ or H-2b+CD8+) isolated from MLNs of recipient mice is depicted. Data are presented as the mean ± SEM and are the cumulative results from 2 experiments (n = 4 mice per group). Statistics: **P ≤ .01, ***P ≤ .001, ****P ≤ .0001.(E) Lethally irradiated (900 cGy) Balb/c mice were transplanted with B6 BM (6 × 106) and purified pan-T cells (0.6 × 106) together with DMSO- or BMS493-treated BM-derived DCs (0.6 × 106) from Balb/c mice. Overall survival is depicted. Data are the cumulative results from 3 experiments (n = 12–14 mice per group).
Figure 5:
Figure 5:. RAR-α signaling is involved in mediating RA’s effect on DCs.
(A) Naïve Balb/cJ mice were irradiated at 900 cGy. RNA was extracted from sort-purified CD11c+ DCs isolated from spleens of these mice 18 hours after irradiation. Gene expression of RAR-α, RAR-β, and RAR-γ was analyzed by real-time q-PCR. Data are normalized for β2-microglobulin RNA and presented as fold increase over gene expression in sort-purified CD11c+ DCs from naïve animals. Data are the cumulative results from two independent experiments.(B, C) Highly purified CD11c+ DCs were isolated from the spleen of naïve Balb/cJ mice. Equal numbers of irradiated (3000 cGy) or non-irradiated DCs at 5 × 104 per well (B) or 1 × 104 per well (C) were co-cultured with 1 × 105 purified spleen pan-T cells labeled with CellTrace Violet dye from naïve B6 animals. Four days later, cells were harvested for flow cytometric analysis. Mean (± SEM) percentage of proliferating alloreactive CD4+ and CD8+ T cells expressing α4β7 from 7 wells is shown. Data are the cumulative results of two experiments.(D, E) Purified CD11c+ DCs were isolated from naïve Balb/cJ mice and incubated with 1 μM BMS195614, LE135, MM11253 (D), or BMS493 (E) for 2–3 hours and washed extensively with cell culture media. 5 × 104 DCs from each treated group were co-cultured with 1 × 105 purified spleen pan-T cells labeled with CellTrace Violet dye from naïve B6 animals. Four days later, cells were harvested for flow cytometric analysis. Mean (± SEM) percentage of alloreactive CD4+ and CD8+ T cells expressing α4β7 from 5 wells is shown. Data are the cumulative results from 2 experiments. Statistics: *P ≤ .05, **P ≤ .01, ****P ≤ .0001.
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References

    1. Shlomchik WD. Graft-versus-host disease. Nat Rev Immunol. 2007;7(5):340–352. - PubMed
    1. Welniak LA, Blazar BR, Murphy WJ. Immunobiology of allogeneic hematopoietic stem cell transplantation. Ann Rev Immunol. 2007;25:139–170. - PubMed
    1. Choi SW, Reddy P. Current and emerging strategies for the prevention of graft-versus-host disease. Nat Rev Clin Oncol. 2014;11(9):536–547. - PMC - PubMed
    1. Zeiser R, Blazar BR. Acute graft-versus-host disease - biologic process, prevention, and therapy. N Engl J Med 2017;377(22):2167–2179. - PMC - PubMed
    1. Shlomchik WD, Couzens MS, Tang CB, et al. Prevention of graft versus host disease by inactivation of host antigen-presenting cells. Science 1999;285(5426):412–415. - PubMed

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