Differentiation of Human T-cell Subsets

The differentiation of human T-cell subsets from naive CD4⁺ T cells was performed as previously described.52 53 (link) Briefly, the treated DCs were purified and co-cultured with human naive CD4⁺ T cells at a ratio of 1:10 in plate-bounded anti-CD3/CD28 (2 µg/mL, Bio X Cell) in normal T-cell culture medium for 6 days. For Th1 differentiation, DCs and naive T cells were co-cultured at a ratio of 1:10 in the presence of anti-IL-4-neutralizing antibody (10 µg/mL, 11B11; Bio X Cell) and recombinant human (rh)IL-12 (10 ng/mL; R&D Systems). For Th17 differentiation, DCs and naive T cells were co-cultured at a ratio of 1:10 in the presence of rhIL-1β (10 ng/mL), rhIL-6 (20 ng/mL), rhIL-23 (10 ng/mL) and recombinant human transforming growth factor-beta (rhTGF-β) (3 ng/mL) (R&D System) for 6 days.