Figure 3.

VitD3 reduced the capability of BMDC1 to induce alloantigen‐specific cytotoxic T lymphocytes (CTL) in a mixed lymphocyte reaction (MLR). Spleen cells of C57BL/6 were stimulated with mytomicin C (MMC)‐treated allogeneic BMDC0 or BMDC1 generated with or without VitD3 for 4 days. (A) The production of interleukin (IL)‐4 and interferon (IFN)‐γ from CD8⁺ T cells stimulated with BMDC0 (a,b) or BMDC1 (c,d) generated in the absence (a,c) or in the presence (b,d) of VitD3 was determined by intracellular cytokine staining. Numbers represent the percentage of cells in each quadrant. The same results were obtained in three separate experiments. (B) After 4 days of MLR, cells were pulsed with ³[H]‐thymidine for 4 h and harvested on a glass filter. Proliferation of spleen cells stimulated with BMDC subsets was determined by counting incorporated ³H using a β‐counter. The bars represent the mean ± SE of triplicate samples. (C) Alloantigen‐specific CTL activity of C57BL/6 mouse spleen cells was measured after 4 day culture with BALB/c BMDC subsets generated in the presence or absence of VitD3. After 4 days of MLR, cells were harvested and their cytotoxicity against P815 mastocytoma cells (H‐2^d) was measured using a 4‐h ⁵¹Cr‐release assay. The bars represent the mean ± SE of triplicate samples. □, BMDC0; ▵, BMDC1; ▪, VD3‐BMDC0; ▴, VD3‐BMDC1.