Figure 2. KOR in immune cells does not contribute to EAE pathogenesis.

(a) Analysis of CD4⁺ T cell subtypes in the spleen of naive, WT-EAE and WT-EAE mice treated with U50488 (1.6 mg kg⁻¹) on day 12 post immunization. Th1, Th17 and Treg cells were analysed with FACS by intracellular staining of IFN-γ, IL-17A and Foxp3, respectively. (b) Analysis of Th1 and Th17 cells in the spleen of naive, WT-EAE or KOR^−/−-EAE mice on day 12 post immunization. (c) In vitro differentiation of Th1, Th17 and Treg cells from WT or KOR^−/− CD4⁺ T cells in the presence of U50488 (10 μM) or not. Data are means±s.e.m. (n=3), *P<0.05, **P<0.01, ***P<0.001 versus naive mice (Student's t-test). (d) EAE scores of Rag1^−/− mice reconstituted with splenocytes (1 × 10⁷) from WT or KOR^−/− mice and immunized with MOG_35–55. (e) EAE scores of lethally irradiated WT mice reconstituted with WT or KOR^−/− bone marrow cells (1 × 10⁷) and immunized with MOG_35–55. (f) EAE scores of lethally irradiated WT or KOR^−/− mice reconstituted with WT bone marrow cells and immunized with MOG_35–55. ^###P<0.001 (two-way analysis of variance (ANOVA) test), *P<0.05, KOR^−/− versus KOR^+/+ recipients (Mann–Whitney U-test). (g) Clinical scores of passive EAE in WT mice induced by transferring MOG-restimulated splenocytes (1 × 10⁷) isolated from WT or KOR^−/− EAE mice. (h) Clinical scores of passive EAE in WT or KOR^−/− mice induced by transferring of MOG-restimulated splenocytes (1 × 10⁷) isolated from WT EAE mice. ^###P<0.001 (two-way ANOVA test), *P<0.05, **P<0.05, KOR^−/− versus KOR^+/+ recipients (Mann–Whitney U-test). Data are means±s.e.m.