Differentiation of human CD4+ T cell subsets in vitro was performed as previously described (69 (link)–72 (link, link, link)). In brief, naive CD4+ CD45RA+ T cells were enriched from donor PBMCs using the T cell enrichment kit (Miltenyi). Cell culture was in the presence of anti-CD3/CD28 activation beads (1:20 bead to cell ratio) (Invitrogen) plus polarization condition medium (RPMI-1640 containing 10% knockout serum replacement [ThermoFisher]) and supplemented with cytokines (vide infra) at 37 °C for 6 d. For Th1 differentiation conditions, naive T cells were cultured in the presence of anti–IL-4 neutralizing antibody (10 mg/mL; R&D) and recombinant human IL-12 (5 ng/mL; R&D). For Th2 conditions, naive T cells were cultured in the presence of anti–IFN-γ neutralizing antibody (10 mg/mL; R&D) in addition to recombinant IL-4 (4 ng/mL; Peprotech). To obtain differentiated Th17 cells, naive T cells were cultured using IL-1β (20 ng/mL; Peprotech), IL-6 (20 ng/mL; Peprotech), recombinant TGF-β (3 ng/mL; Peprotech), and IL-23 (10 ng/mL; R & D) for 7 d. To expand Th17 cells, Dynabeads (Invitrogen) isolated CD4+ T cells were used instead of CD45RA+ selected T cells.