Increasing evidence indicates the existence of membrane receptors for testosterone (mAR) and estradiol (mER) on the surface of cells, besides the classic intracellular androgen receptor (iAR) and estrogen receptors (iER). Here, we investigate the occurrence of sex steroid receptors in B cells isolated from the spleen of C57BL/10 mice using magnetic cell sorting. RT-PCR reveals the presence of iAR, iERalpha, but not iERbeta. Using different anti-iAR and anti-iER antibodies flow cytometry and confocal laser scanning microscopy (CLSM) localize iAR and iERalpha in the cytoplasm, which are translocatable to the nucleus upon incubation with testosterone (T) and 17beta-estradiol (E(2)). The surface of B cells is devoid of iAR and iERalpha and does not bind any T and E(2) conjugated to BSA-FITC as revealed by flow cytometry and CLSM. In accordance, T and E(2) are not able to induce any rapid rise in in the intracellular free Ca2+ concentration of Fura-2 loaded B cells. Our data indicate that B cells express neither mAR nor mER on their surfaces, in contrast to other major cells of the immune system such as T cells and macrophages.