Figure 7. Th cell–specific Prmt5 deficiency prevents induction of EAE autoimmunity.

(A) Schematic of tamoxifen treatment/EAE experimental design and downstream analyses. (B) EAE score in iCD4-PRMT5^Δ/Δ and indicated controls after MOG_35–55/CFA immunization. All mice were treated with tamoxifen by oral gavage before immunization. (C–E) Flow analysis and quantification of CNS-infiltrating (C) CD3⁺ and CD3⁺CD4⁺ T cells or (D and E) naive, Tem, and Tcm phenotype CD4⁺ T cells from iCD4-PRMT5^Δ/Δ and indicated controls 21 days after MOG_35–55/CFA immunization. (F–Q) CNS-infiltrating cells (F–K) or splenocytes (L–Q) from day 14 iCD4-PRMT5^Δ/Δ and indicated controls were reactivated with MOG_35–55. (F and L) Proliferation was monitored by ³H-thymidine incorporation and expressed as a relative proliferation ratio to the resting PRMT5^fl/fl media control condition. MOG_35–55-reactivated cells were analyzed by ELISA for (G and M) IFN-γ and (H and N) IL-17 secretion, and flow cytometry for (I and O) T-bet⁺IFN-γ⁺ Th1, (J and P) RORγt⁺IL-17⁺ Th17, and (K and Q) T-bet⁺IL-17⁺ cell populations. Data are pooled from 4 independent experiments, n = 6–10 mice. Kruskal-Wallis with Dunn’s multiple-comparisons test (B and Q); 1-way ANOVA followed by Sidak’s multiple-comparisons test (F, L, O, and P) or Student’s t test (C, E, G–K, M, and N). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Box-and-whisker plots boxes extend from 25th to 75th percentiles, all points shown, whiskers extend from min to max, line represents median. Bar graphs are indicated as mean ± SD for F, I–L, O–Q and mean ± SEM for G, H, M, and N. Tem, effector memory T cells; Tcm, central memory T cells; TF, transcription factor; Ck, cytokine.