FIGURE 4.

Anti-TNFR2 antibody reduces the proliferation and suppressive activity of regulatory T cells (Tregs). CD4⁺CD25^– T cells (Teffs) and CD4⁺Foxp3⁺ T cells (Tregs) from pooled spleen and lymph nodes (inguinal, axillary, and mesenteric regions) of BALB/c mice were purified. (A) Carboxyfluorescein succinimidyl ester (CFSE)-labeled CD4⁺Foxp3⁺ T cells (Tregs) were treated with anti-TNFR2 antibody (1 and 10 μg/ml) or isotype IgG (10 μg/ml) for 72 h. CFSE dilution analysis showed that anti-TNFR2 antibody inhibited the proliferation of Tregs. (B) Summary of the percentage of proliferating CD4⁺Foxp3⁺ Tregs with CFSE dilution from three independent experiments. (C) Tregs pretreated with anti-TNFR2 antibody (10 μg/ml) or isotype IgG (10 μg/ml) were cocultured with CFSE-labeled CD4⁺CD25^– Teffs at different ratios in the presence of irradiated APCs and 5 μg/ml of anti-CD3 antibody for 72 h. Anti-TNFR2 antibody pretreatment reduced the suppressive activity of Tregs. Data are the summary of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.