Figure 8. Bid KO cDCs promote increased OT-2 cell expansion and function in lymph node and spleen.

cDCs from topically immunized ear skin explants were injected (3 × 10⁵) into the footpad of WT recipient mice (3 per group) containing CD45.1 allotypic OT-2 T cells (2 × 10⁶). Control mice were topically immunized on ears (see Materials & Methods). A. Lymph node cells were harvested after 4 days and stained for CD45.1 to reveal OT-2 T cells (upper panel) that were gated for analysis of cytokine staining (lower panel). B. Spleen cells harvested after 4 days were restimulated with 100 μg/ml OVA in vitro for 3 days prior to CD45.1 and cytokine staining. C. Lymph node weights are increased in mice receiving Bid KO cDCs. Lymph nodes from mice receiving PBS (open bars) or immunization by topical OVA or by cDC transfer (filled bars) were weighed and the mean ± SD per group calculated. *, P < 0.05. D. and E. Bid KO cDCs promote increased numbers of IFNγ producing cells in lymph node (D) and spleen (E). The number of IL-4 (open bar) or IFNγ (filled bar) producing OT-2 cells per million CD3 T cells was calculated for each individual mouse and mean ± SD determined per group. *, P < 0.05. Representative density plots and statistical analyses are shown for one of three independent experiments.