Figure 7

Effects of induced antigen-specific regulatory T cell transfer on T-cell infiltration in the brain tissue in the amyloid-β42-induced autoimmune encephalomyelitis model. CD4⁺CD25⁻Foxp3⁺ induced antigen-specific regulatory T cells (iTregs) were isolated from coimmunized Foxp3 enhanced green fluorescent protein (Foxp3-eGFP) mice on day 7 after the second coimmunization and adoptively transferred into amyloid-42β-induced autoimmune encephalomyelitis (Aβ42-EAE) model mice on days 5 and 12 after EAE induction. (A) Schematic representation of iTreg cell transfer to prevent Aβ42-EAE. iTregs were isolated from Foxp3-eGFP mice after the second coimmunization and adoptively transferred into Aβ42-EAE model mice on days 5 and 12 after the model was induced. CD4⁺CD25⁺Foxp3⁺nTreg transfers were used as a control. CFA, Complete Freund’s adjuvant; nTreg, näive regulatory T cell; PT, Pertussis toxin. (B) Single-cell suspensions of brains were isolated from EAE-induced animals with or without vaccinations, or from naïve C57BL/6 mice, and stained on day 21. The cells were immunostained for CD3 and CD4 and analyzed by fluorescence-activated cell sorting. MOG, Myelin oligodendrocyte glycoprotein. (C) The number of double-positive CD3⁺ and CD4⁺ T cells per 10⁶ brain cells taken from the groups of six animals detected the same way as in (B) were statistically analyzed and plotted. The results shown are typical of three independent experiments. All data are presented as mean ± SD. Statistical analysis was performed using parametric one-way analysis of variance, and t-tests were used to compare two groups. ***P < 0.001 compared with Aβ42-EAE mice.