FIGURE 5.

RelB^ΔDC mice are protected from EAE due to accumulation of Tregs EAE was induced by immunizing littermate control or RelB^ΔDC mice with MOG_35–55 in CFA as described in Materials and Methods. (A) EAE clinical score in WT and RelB^ΔDC mice shown as mean ± SEM of one representative experiment. Table depicts summary of all experiments. Sick, score of at least one. Mice that had to be sacrificed because of high disease scores are indicated as deaths. (B) Total numbers of cytokine-expressing Th cells in the CNS at peak of disease after PMA/ionomycin restimulation of one representative experiment. (C) Frequency of Foxp3⁻Gata3⁺ and Foxp3⁺Gata3⁺ Th cells in the CNS at peak of disease. (D) Frequency of Foxp3⁺ cells within CD4⁺ T cells or frequency of Helios⁺ ST2⁺ cells within Foxp3⁺ Tregs in the CNS at peak of disease. (E) Clinical score of EAE in littermate control and RelB^ΔDC mice treated with an anti-CD25 Ab (open symbols) or PBS (filled symbols) prior induction of EAE as described in Materials and Methods section. Diagram shows mean ± SEM. (F) Total cell number of cytokine-expressing Foxp3⁻ CD4⁺ T cells in the CNS at the peak of EAE. (G) Littermate control and RelB^ΔDC mice received 2.5 × 10⁶ sort-purified MOG-specific Foxp3/GFP⁻2D2 T cells prior EAE induction via i.v. injection. Upper plots (left) and quantification (right) indicate frequency of Vα3.2⁺Vβ11⁺ 2D2 cells among CD4⁺ T cells isolated from draining inguinal lymph nodes at day 8 after EAE induction. Lower plots indicate frequency (left) and quantification (right) of Foxp3/GFP⁺ cells among Vα3.2⁺Vβ11⁺ 2D2 T cells isolated from draining inguinal lymph nodes at day 8 after EAE induction. Bar diagrams show mean ± SD of at least two independent experiments unless otherwise indicated. Statistical analysis was performed using Mann–Whitney U test (in [A]) or two-way ANOVA with Tukey multiple correction test in (E). Otherwise, unpaired two-tailed Student t test was used. *p < 0.05, **p < 0.01, ***p < 0.001.