Fig. 3

TLR4^−/− CD4⁺ naïve T cells are defective in promoting EAE. Rag1^−/− mice transplanted with wild type (WT) or TLR4^−/− CD4⁺ naïve T cells are immunized with MOG for EAE induction. a Mean EAE scores of mice (n = 5 per group). b–b” H&E staining (b), LFB staining (b’), and MOG immunofluorescence staining (b”) of spinal cord sections from transplanted Rag1^−/− mice. Arrows show lymphocyte infiltration. Scale bar, 300 μm. Quantification analyses are on the right. c Western blot and quantification analyses of MOG expression in spinal cord. d Quantification analyses of IL-17⁺ cells among CD4⁺ gated cells from peripheral blood (PB) and spinal cord-infiltrated (SCI) lymphocytes by flow cytometry. e ELISA of IL-17A, IL-17F, and GM-CSF in peripheral blood serum. f Quantitative PCR analyses of Rorγt expression in spinal cord-infiltrated lymphocytes. g, h Splenocytes and lymph node cells from mice presented (a) on day 20 are stimulated in vitro. Flow cytometric analyses of IL-17⁺ and GM-CSF⁺ cells among CD4⁺ gated cells (g). The concentrations of IL-17A, IL-17F, and GM-CSF in supernatant are measured by ELISA (h). Data are presented as mean ± standard deviation. *P < 0.05. **P < 0.01. ***P < 0.001; two-way repeated measures ANOVA (a), Student’s t test (b–h). Data are representative of three experiments done in triplicate