Fig. 2
From: K27-linked RORγt ubiquitination by Nedd4 potentiates Th17-mediated autoimmunity
Nedd4 Expression in T cells confers the susceptibility of mice to EAE induction and is required for antigen-specific Th17 response. EAE was induced by MOG35-55 in CFA, and the disease activity was monitored for 21–25 days after immunization. For ex vivo T cell responses, Cd4 Cre and Cd4 Cre-Nedd4f/f mice, or Tcrb–/– mice adoptively transferred with naïve CD4+ T cells from Nedd4f/f and Rorc Cre-Nedd4f/f mice were immunized with MOG35-55 in CFA, and mice were analyzed at day 8 after immunization for Th1/Th17 in draining lymph nodes (dLNs) and at day 14–16 for total infiltrating cells and Th1/Th17 in the spinal cords. A EAE scores of Nedd4CreER mice pretreated with tamoxifen or corn oil, or B6 mice (n = 5 mice per group) pretreated with or without tamoxifen. WT, WT-TAM, or Nedd4CreER−oil vs. Nedd4CreER−TAM. *p < 0.05, **p < 0.01; Mann–Whitney U test. B EAE scores of Nedd4f/f (n = 6), Cd11c Cre-Nedd4f/f (n = 5), LysM Cre-Nedd4f/f (n = 6) mice. C EAE scores of Cd19 Cre (n = 7) and Cd19 Cre-Nedd4f/f (n = 6) mice. D EAE scores of Cd4 Cre (n = 8) and Cd4 Cre-Nedd4f/f (n = 5) mice. *p < 0.05, **p < 0.01; Mann–Whitney U test. E Flow cytometric analysis of CD4+IFN-γ+ Th1 cells and CD4+IL-17+ Th17 cells of Cd4 Cre and Cd4 Cre-Nedd4f/f mice (n = 3 per group) at day 8 after immunization. **p < 0.01; student t test. F Flow cytometric analysis of I-Ab-MOG35-55+IL-17+ T cells of Cd4 Cre and Cd4 Cre-Nedd4f/f mice (n = 3 mice per group) at day 8 after immunization. *p < 0.05; student t test. G Numbers of infiltrating cells in CNS from Cd4 Cre and Cd4 Cre-Nedd4f/f mice (n = 3 mice per group) at day 14–16 after immunization. ***p < 0.001, student t test. H Flow cytometric analysis of CD4+IFN-γ+ Th1 cells and CD4+IL-17+ Th17 cells in spinal cords of Cd4 Cre and Cd4 Cre-Nedd4f/f mice (n = 3 per group). *p < 0.05, student t test. I EAE scores of Tcrb–/– mice (n = 7 mice per group) adoptively transferred with CD4+ T cells from Nedd4f/f and Rorc Cre-Nedd4f/f mice upon immunization with MOG35-55 in CFA. *p < 0.05, **p < 0.01; Mann–Whitney U test. J Flow cytometric analysis of Th1/Th17 cells in the dLNs of Tcrb–/– mice (n = 3 mice per group) received CD4+ T cells (5 × 106) from Nedd4f/f and Rorc Cre-Nedd4f/f mice at day 8 after immunization. Data are representative of two to three independent experiments