FIGURE 9.

CD25-IL2 FP augmented the percentages of FOXP3⁺ Tregs in vivo. On days 0 and 14, 2D2-FIG mice (n = 4/group) were vaccinated with saline, CD25-IL2 alone, or the combination of CD25-IL2 and MOG35-55. All injections were administered subcutaneously in Alum. Vaccinations included 3 or 2 nmoles FP (day 0 and 14, respectively) and 4 nmoles MOG35-55. PBMCs were assayed for CD3, CD4, GFP (FOXP3), CD25, and Vβ11 by flow cytometry on days 4, 10, and 18. Shown are (A) representative dot plots (day 18 time-point) from each treatment group gated on CD3⁺ CD4⁺ T cells and analyzed for CD25 and FOXP3 expression, (B) percentages of FOXP3⁺ Tregs (CD3⁺ CD4⁺ T cell gate) before vaccination (day 0) and for the day 4, 10, and 18 time points, (C) the FOXP3 MFIs (CD3⁺ CD4⁺ FOXP3⁺ Treg gate), and (D) the CD25 MFIs on Tregs (left, CD3⁺ CD4⁺ FOXP3⁺ gate) and Tcons (right, CD3⁺ CD4⁺ FOXP3^– gate). (D) Note the use of different y-axis scales. Statistical significance was analyzed by a one-way ANOVA with the Holm-Sidak multiple comparisons test. Statistically significant (p < 0.05) differences were noted as indicated (a, 1 vs 2) (b, 1 vs 3). Error bars represent SEM.