Figure 2

EAE in myeloid cell-specific Il27p28^−/− mice. LysM^WT (n = 7) and LysM^Cre Il27p28^fl/fl (LysM^ΔIl27p28) (n = 11) mice were induced for EAE. (a) EAE clinical score. (b,c) The numbers of CNS infiltrating CD4⁺ and CD4⁺Foxp3⁺ Treg cells, and the mean fluorescence intensity (MFI) of Foxp3 was determined by flow cytometry at day 17 post immunization. (d) At day 18 postimmunization, tissue sections of the spinal cords were stained with H&E. Arrows indicate infiltration of inflammatory cells. × 200 magnification. (e) Flow cytometry analysis of GM-CSF, IFNγ, IL-17, and TNFα CD4⁺ T cells from the CNS of EAE mice (day 17 post immunization). (f) The levels of IL-6, IFNγ, TNFα and IL-17A in the serum of EAE mice (day 17 post immunization) were measured using Cytometric Bead Array. Each serum sample was analyzed in duplicates. (g) qPCR analysis of the indicated mRNAs in the brain and spinal cords from naïve, LysM^WT, and LysM^ΔIl27p28 mice 17 days post immunization. Gene expression was normalized by Gapdh and compared to that of naïve mice. n = 3 per group. (h) qPCR analysis of the indicated mRNAs in freshly sorted CD45^high CD11b^high (infiltrating myeloid cells), CD45^int CD11b^high (microglia) and CD45^low (astrocyte and oligodendrocyte) cells from LysM^WT or LysM^ΔIl27p28 mice 17 days post immunization. n = 3 per group. *p < 0.05; **p < 0.01; ***p < 0.001; as determined by Mann–Whitney nonparametric test.