Figure 6.

IL-17 production by biTregs is dependent on RORγt. (A) Representative FACS plots of spleen cells 6 days after sheep IgG immunization of indicated mice (pre-gated on CD45⁺CD3⁺CD4⁺ events). Quantification of RORγt in CD4⁺ and Foxp3⁺ T cells in Foxp3^Cre (n=11) and RORC^flox/flox × Foxp3^Cre mice (n=15). (B) Quantification of spleen cell numbers. (C) Splenic CD4⁺ subsets determined by FACS (gated as indicated). (D) IL-17 production by splenic Foxp3⁺ T cells. (E) T cell activation and (F) proliferation of the indicated subsets as determined by flow cytometry from spleen cells. (G) Spleen cell secretion of the indicated cytokines determined by ELISA. (H) In vitro suppression of T helper cell (Th) produced IL-2 and IFNγ by WT and RORγt^−/− Tregs as determined by ELISA. Error bars represent standard deviation. ***P<0.001.