Assays to analyze the cytokines involved in DC maturation. A-E, ELISA assay for IFN-γ (A), IL-12 (B), IL-6 (C), IL-10 (D) and TGF-β (E) using the culture supernatants of imDCs, mDCs (TNF-α-treated DCs) and TNF-α with PsL-EGFmAb-treated DCs in vitro. F, Neutralization assays. PsL-EGFmAb-treated DCs were co-cultured with freshly isolated CD4+CD25+ Tregs only (control) or in the presence of one of the following neutralizing anti-human antibodies in vitro: anti-IL-6, anti-IL-10, anti-TGF-β, anti-IL-4, anti-TNF-α, anti-IL-12 or anti-IFN-γ mAbs. The isotype-matched controls were also used. Allogeneic mixed T cell proliferation assays were performed as above. The results are shown as fold increases in [3H]TdR incorporation. G, Neutralization assays. CD4+CD25+ T cells were co-cultured with PsL-EGFmAb-treated mDCs (T1 cell), then mixed with CD4+CD25- effector T cells. Suppression assays were performed in the presence of the neutralizing antibodies mentioned above or with medium only (control). The mean ± SD of three independent experiments is shown. *p < 0.05, **p < 0.01 vs. control; #p < 0.05, ##p < 0.01 vs. mDC group; +p < 0.05, ++p < 0.01 vs. imDC group.