Induction of T-cell proliferation and IFN-γ secretion by immature and mature MoDCs. Immature MoDCs (GM-CSF plus IL-4) were stimulated for 2 days with the indicated stimuli and used as stimulators (3 × 10⁴) of CFSE-labeled CD3⁺ alloreactive T cells (3 × 10⁵) in the MLR. On day 5 of the MLR, T cells were restimulated with the identically conditioned MoDCs for 8 hours in the presence of Brefeldin A, and CD4⁺ T cells were assessed for intracellular IFN-γ secretion and proliferation by FACS analysis by gating on CD3⁺CD8^– T cells. Percentages indicate the percent IFN-γ⁺CD4⁺ T cells within the gated region. Data are representative of experiments from 7 separate donors. Similar profiles were observed when CD3⁺CD8⁺ T cells were examined.