Figure 4

Induction of Foxp3⁺ Tregs by tDCs. (a) Generation of Foxp3⁺ Tregs from CD4⁺CD25⁻ OT-II T cells. Mature cDCs, mDCs, dDCs, or mdDCs generated from BM cells of C57BL/6 mice were pulsed with the OVA_323–339 peptide (10 μg/ml), washed, and co-cultured with CD4⁺CD25⁻ T cells isolated from OT-II mice in the presence of 100 U/ml IL-2. After 4 days, cells were analysed for the expression of CD25 and Foxp3. Representative histograms are shown. (b) The proportion of CD25⁺Foxp3⁺ T cells in the CD4⁺ cell population of each experimental group of (a) is shown. The data present the mean ± SD of three independent experiments. (c) Generation of Foxp3⁺ Tregs in allogenic MLR. Mature cDCs, mDCs, dDCs, or mdDCs generated from BM cells of C57BL/6 mice were cultured with naïve CD4⁺CD25⁻ T cells isolated from the spleens of BALB/c mice for 4 days in a medium containing 10 U/ml of recombinant human IL-2. After 4 days, cells were analysed for the expression of CD25 and Foxp3. The proportion of CD25⁺Foxp3⁺ T cells in the CD4⁺ cell population of each experimental group is shown. The data present the mean ± SD of three independent experiments. (d) Effects of anti-IL-10 neutralizing monoclonal antibody on the generation of Foxp3⁺ Tregs from CD4⁺CD25⁻ OT-II T cells. In the experimental system of (a), anti-IL-10 monoclonal antibody, or isotype-matched monoclonal antibody (10 μg/ml) was added from the initiation of the culture. The data present the mean ± SD of three independent experiments. One way ANOVA tests were performed in order to evaluate significance. ^#P < 0.05, ^##P < 0.01 compared with cDC group. *P < 0.05, **P < 0.01 compared with matched group.