Figure 2. SIGIRR deficiency leads to increased susceptibility of Th17-dependent EAE and hyper activation of MOG-specific Th17 cells.

(A) EAE was induced by MOG_35–55 immunization. Mean clinical scores were calculated each day for WT (n = 13) and Sigirr^−/− mice (n = 10). (B) Hematoxylin and eosin and anti-CD3 staining of spinal cord of WT and Sigirr^−/−mice 15 days after immunization with MOG_35–55. (C) Immune cell infiltration in the brain of MOG_35–55 immunized WT and Sigirr^−/− mice (n=3, 7 days after disease onset) was analyzed by flow cytometry. (D) Real-time PCR analysis of relative expression of IFN-γ, IL-17, TNF-α and IL-6 in spinal cords of MOG_35–55 immunized WT and Sigirr^−/− mice (n=3, 7 days after disease onset) as compared to CFA treated WT control mice. Error bars, s.d.; *, p<0.05; **.p<0.01 (two tailed t-test). Data are representative of three independent experiments. (E–F) Draining lymph node cells from wild-type mice and Sigirr^−/− mice were collected 10 days after immunization with either MOG_35–55 emulsified in complete Freund's adjuvant or complete Freund's adjuvant alone and were re-stimulated with MOG_35–55 in vitro for 4 days, followed by ELISPOT analysis (E) and ELISA (F) of IL-17 and IFN-γ. Error bars, s.d.; n = 10 mice per group. p<0.05; **.p<0.01 (two tailed t-test). (G) Primed MOG_35–55 specific T cells (10 days) were re-stimulated with MOG_35–55 in vitro in the presence of recombinant IL-23 for 4 days, and then transferred to naïve wild-type and Sigirr^−/− mice. Graph represents the average clinical score after T-cell transfer. n=5. *, p<0.05; (ANOVA). Data are representative of three independent experiments.