Figure 5
Loss of IL-17F does not impact on T cell priming and does not lead to compensatory upregulation of IL-17A.
(A–C) Mice were immunized with MOG35–55/CFA and lymphocytes were isolated from LNs prior to disease onset at 7 days after immunization. Cells were rechallenged with 50 μg/ml of MOG35–55, and IL-17A and IL-2 were measured by ELISA (A) and ELISPOT (B). (C) Proliferation of effector Th cells upon stimulation with MOG35–55 peptide or concanavalin A (ConA) was measured by thymidine incorporation. (A–C) A representative of 3 independent experiments is shown. Error bars indicate SEM of measured replicates. (D) Splenocytes from naive mice were polarized toward the Th17 effector type in vitro, and IL-17A, IL-17F, and IFN-γ were measured by intracellular cytokine staining. Dot plots are gated on Th cells (CD4+), and histograms are gated on Th17 cells (CD4+, IL-17A+). (E) Within the IL-17A–expressing Th17 compartment, IL-17F production is shown for each genotype. (F) CNS-infiltrating lymphocytes were isolated on day 24, after immunization from severely sick mice, restimulated with PMA/ionomycin and Brefeldin A for 5 hours, and analyzed for their IL-17A and IFN-γ expression by flow cytometry. (D–F) Percentages of gated cells are shown.
