Figure 4. In Vivo Inhibition of Autophagy Induces CD4⁺ T Cell Tolerance and Decreases the Severity of MOG-Induced EAE.

(A–C) Wild-type or NFAT1-deficient OT-II (CD90.2⁺) T cells were transferred into congenic C57BL/6 (CD90.1⁺) mice (A). 24 hours later, mice were challenged with subcutaneous OVA_323–339 in CFA. For each group, half of the mice were randomly treated with chloroquine and half with PBS for 6 days. IL-2 production (B) and Grail and Egr2 expression (C) were determined in purified OT-II T cells from the draining lymph nodes and stimulated ex vivo with T cell-depleted splenocytes loaded with OVA_323–339.

(D) Atg7 content and autophagy flux (LC3-II turnover) measured by immunoblot in resting and activated CD4⁺ T cells isolated from Atg7^−/− or wild-type mice.

(E) EAE scores of Atg7^−/− or wild-type littermate mice immunized with MOG_35–55 peptide.

(F) Histological analyses of lumbar spinal cord sections from EAE Atg7^−/− or wild-type mice.

(G) EAE was induced in C57BL/6 mice that were divided in two groups that received daily injections of PBS or chloroquine in PBS (Chloro). EAE scores were recorded daily.

(H) Histological analyses of lumbar spinal cord sections from untreated and chloroquine-treated EAE mice.

(I and J) Experiments were performed as in (G) and (H) but using NFAT1-deficent mice.

Data represent mean + SEM from four different experiments (B and C). *p < 0.05; **p < 0.01; ns, not significant (ANOVA with Tukey post-test in B and two-tailed t test in C). For (E)–(G), data represent mean and SEM from 8 (E), 10 (G), or 4 (I) different mice from 2 independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 (Mann-Whitney test). See also Figure S4.