Modulation of human lymphocyte marker expression by gamma irradiation and mitomycin C

gamma irradiation (GR) or mitomycin C (MC) treatment of stimulator cells is frequently used to achieve unidirectionality of response in the mixed lymphocyte reaction. As GR differs from MC in the pathways used to block lymphocyte replication, this study analyzes the effects of these modalities upon the expression of various differentiation and Class II major histocompatibility antigens on lymphocytes cultured for 24, 48, and 72 hr. There was a decrease in the mean density of HLA-DR expression on CD3+ and on CD8+ cells at 24, 48, and 72 hr after exposure to GR (42 and 35, 60 and 69, and 26 and 49%, respectively) or to MC (26 and 11, 26 and 18, and 46 and 30%, respectively). There was a parallel decrease in the levels of the corresponding cell subsets when compared with control cultured cells not exposed to GR or MC. In contrast, the density of HLA-DR markers on CD3-negative cells was increased at 24, 48, and 72 hr of culture following exposure to GR (73, 82, and 102%, respectively) or to MC (9, 45, and 80%, respectively). There was a more profound decrease in CD3+, CD8+, and CD19+ cell subset levels and in the density of the corresponding markers in GR-treated cells than in those of cells exposed to MC when the results were compared with those of untreated cultured control cells. Although GR appears to exert a more profound effect than MC, the results indicate that both modalities have the capacity to reduce the density of polymorphic determinants on Class II (HLA-D region)-encoded molecules on T (CD3+ and CD8+) and B (CD19+) cells which are known to trigger potent MLR responses. Both modalities may therefore affect profoundly the relative strength of MLC responses and the derived measurements of the degree of HLA Class II compatibility between stimulator and responder cells.