Fig. 3: Loss of Treg cells in the postinflammatory CNS rekindles inflammation.
a, EAE was induced in Foxp3 (GFP) and Foxp3-DTR mice. DTx was administered i.c.v. or i.p. during the recovery phase of EAE. Mice were killed 3 days after DTx injection. Brains were collected for immunohistochemistry staining. Arrows and squares indicate immune cell niches. Representative stainings for CD45, CD4 and IBA-1 are shown; scale bars, top row: 800 μm, bottom three rows: 50 μm. b, Quantification of CD45+ and CD4+ cells as percentages and absolute counts per mm2. Symbols represent individual mice. Data are shown as mean + s.d. P values were calculated using a one-way ANOVA with a Tukey’s HSD test (DTx i.c.v. control n = 3, DTx i.p. n = 5, DTx i.c.v. n = 9 biological replicates). c,d, EAE was induced in Foxp3-DTR mice, and DTx was administered i.c.v. during the recovery phase. FTY720 (FTY; 1 μg per gram per day) or PBS was administered i.p. from 24 h before to 72 h after DTx i.c.v. injection. CD4+ T cell counts in the CNS (c) and MFI of CD69 on CNS CD4+ Tconv cells (d) as determined by flow cytometry are shown. Symbols represent individual mice. Data are shown as mean + s.d. The P value in c was determined by a two-tailed unpaired Student’s t-test (DTx i.c.v. plus PBS n = 4, DTx i.c.v. plus FTY n = 3 biological replicates). e, EAE scores after i.c.v. DTx administration shown as mean + s.e.m. (DTx i.c.v. plus PBS n = 4, DTx i.c.v. plus FTY n = 4 biological replicates).
