Figure 7.

Gp96 activates CD4⁺Foxp3⁺ Tregs through MyD88 signal

(A–C) Foxp3^CreMyd88^fl/fl and Foxp3^CreMyd88^+/+ mice were treated as in Figure 6B. OVA-specific IgG levels were determined in serum on day 20 post OVA challenge using ELISA (A). FACS analysis of CXCR5⁺PD-1⁺ cells (Tfr) (B) in spleen and CD4⁺Foxp3⁺ Tregs (C) in the indicated organs.

(D) FACS analysis of Foxp3, Helios, CD62L, CD69, ICOS and CD137 expression on CD4⁺Foxp3⁺ Tregs in spleens of Foxp3^CreMyd88^fl/fl and Foxp3^CreMyd88^+/+ mice immunized with 200 μg gp96 or saline (control).

(E) Flow cytometry analysis of ICOS, KLRG1, CD69 and CD137 expression in CD4⁺Foxp3⁺ Tregs from CD45.1⁺ cells and CD45.2⁺ cells.

(F) Foxp3^CreMyd88^fl/fl and Foxp3^CreMyd88^+/+ mice were immunized with gp96 or saline (control) before induction of EAE. Disease score was measured daily after EAE induction. Dots represent data from individual mice. The data are representative of two independent experiments with similar results. Mean ± SD is shown. The Student's t test or two-way ANOVA was used for statistical analysis. P < 0.05 were considered statistically significant. ns = not significant.