Figure 2.

Let-7 miRNAs negatively regulate CD4⁺ T cell pathogenicity in a cell-intrinsic manner in EAE. (A) Mean clinical scores in Rag2KO recipient mice that received 2D2Rag2KO WT (n = 7), 2D2Rag2KO Let-7Tg (n = 7) or 2D2Rag2KO Lin28Tg (n = 8) naïve CD4⁺ T cells (2–2.5 × 10⁶ cells/recipient) and that were subsequently immunized with MOG₃₅₋₅₅ in complete Freund's adjuvant (CFA) and pertussis toxin (60 ng). (B) Number of total mononuclear cells at the peak of the disease (day 9 post-immunization) in the CNS of Rag2KO recipients that received 2D2Rag2KO WT, 2D2Rag2KO Let-7Tg, and 2D2Rag2KO Lin28Tg cells. (C) Number of CNS-infiltrated 2D2Rag2KO CD4+ T cells at the peak of the disease (day 9 post-immunization) in Rag2KO recipients transferred with 2D2Rag2KO WT, 2D2Rag2KO Let-7Tg, and 2D2Rag2KO Lin28Tg cells as analyzed by flow cytometry. (D) Intracellular staining of donor CD4⁺ T cells from the CNS of Rag2KO recipients that received 2D2Rag2KO WT, 2D2Rag2KO Let-7Tg, and 2D2Rag2KO Lin28Tg cells (left). Numbers indicate the frequencies of cytokine-positive cells within the indicated gates. **p < 0.01, ***p < 0.001, ****p < 0.0001 (A–C), compared with WT employing two-way ANOVA (A) or using two-tailed Student's t-test (B,C). Data are from two combined independent experiments (A–C; mean ± S.E.M. of each population from all mice) or from one experiment representative of two independent experiments (D).