Immune tolerance was restored in treated EAE mice.

<p>A. At day 4 after the second treatment, T cell proliferation by MTT method was performed with MOG_35–55 peptide as stimulation antigen, ConA as positive control and BSA as irrelevant antigen. B. At day 4 after the second treatment, the splenocytes of treated EAE mice were prepared for Treg cells analysis. Gating on CD4⁺ T cells, Treg cells (CD4⁺CD25⁺Foxp3⁺) were counted relatively to total CD4 cells by flow cytometry. C. Treg cells from naïve mice (nTreg) or FK506/p2MOG₃₅ treated mice (iTreg) were co-culture with CD4⁺CD25⁻ T cells from naïve C57BL/6 mice respectively. Proliferation was tested by MTT method. Bar, mean and SD from 3 independent experiments, each using at least three mice per group (n = 3); *, p<0.05 between the indicated pairs.