Figure 6.
IL-7Rα Expression Promotes Th17 Cell Differentiation
(A) IL-2 signaling inhibits IL-17 production by CD4 T cells activated under Th17-skewing conditions. Naive CD4 T cells enriched from WT mice were stimulated with 1 μg/mL plate-bound anti-CD3/CD28 for 5 days in the presence of 5 ng/mL TGF-β, 30 ng/mL IL-6, 10 μg/mL anti-IL-4, and 10 μg/mL anti-IFN-γ and the indicated concentration of IL-2. Levels of IL-17A and IFN-γ production were assessed by flow cytometry following re-stimulation with PMA/ionomycin in the presence of brefeldin A. Results are the summary of three independent experiments, showing mean with SEM.
(B) Loss of IL-7Rα expression reduces IL-17 production in CD4SP thymocytes. TCRβhiCD4SP thymocytes from WT and IL-7Rα-cKO mice were sorted and stimulated with 1 μg/mL plate-bound anti-CD3/CD28 for 5 days in the presence of 5 ng/mL TGF-β, 30 ng/mL IL-6, 10 μg/mL anti-IL-4, and 10 μg/mL anti-IFN-γ. IL-17A and IFN-γ production was assessed by flow cytometry following re-stimulation with PMA/ionomycin in the presence of brefeldin A. Results are based on three independent experiments with a total of three WT and three IL-7Rα-cKO mice, showing mean with SEM. Unpaired two-tailed Student's t test, where ∗∗p < 0.01.
(C) IL-7Rα overexpression increases IL-17 production in CD4+ LN T cells. Naive CD4+ LN T cells were enriched by magnetic selection and stimulated with 1 μg/mL plate-bound αCD3/CD28 for 5 days in the presence of 5 ng/mL TGF-β, 30 ng/mL IL-6, 10 μg/mL anti-IL-4, and 10 μg/mL anti-IFN-γ. IL-17A and IFN-γ production was assessed by flow cytometry following re-stimulation with PMA/ionomycin in the presence of brefeldin A, showing mean with SEM.
Unpaired two-tailed Student's t test, where ∗p < 0.05. See also Figure S7.