Fig. 4

GMCSF-MOG induced robust Treg responses even when mixed with an immunogenic vaccine. a–n On day 0, 2D2-FIG (n = 3–5) mice were injected with 4 nmol of GMCSF-MOG, 4 nmol of GMCSF-NFM, or saline. Separate groups were also injected with either 2 nmol of GMCSF-MOG + 2 nmol of GMCSF-NFM (a, b) or 4 nmol of GMCSF-MOG + 4 nmol GMCSF-NFM (c–n). PBMCs were assayed for CD3, CD4, FOXP3, Vβ11 (2D2 TCRβ), CD44, and CD62L expression. Shown are a, c percentages and b, d numbers (per microliter of blood) of FOXP3⁺ Tregs for CD3⁺ CD4⁺ T cells collected on days 0, 5, 12, and 19 (a, b) or days 0, 7, and 15 (c, d). Also shown for day 7 are e representative dot plots of CD3⁺ CD4⁺ T cells analyzed for Vβ11 (y-axis) and FOXP3 expression (x-axis); f the total number of CD3⁺ T cells per μl of blood; representative histograms for g Vβ11, i CD3, k CD44, and m CD62L expression of CD3⁺ CD4⁺ T cells; and the mean florescence intensity (MFI) of h Vβ11, j CD3, l CD44, and n CD62L. a, b Statistical significance was analyzed by use of a two-way repeated measures ANOVA. Means for groups G-MOG, “G-MOG + G-NFM”, and G-NFM represented statistically significant differences compared to each other (*p < 0.05). c–n Statistical significance was analyzed by use of a one-way ANOVA; c, d (a) G-MOG vs G-NFM (p < 0.05) and (b) “G-MOG + G-NFM” vs G-NFM (p < 0.05). (*p < 0.05, **p < 0.01, ***p < 0.001). Error bars represent SEM