Fig. 2

The high-efficiency GMCSF-NFM vaccine induced memory Tcon responses in 2D2-FIG mice. On day 0, 2D2-FIG mice were SC injected with 4 nmol of GMCSF-MOG, 4 nmol of GMCSF-NFM, or saline. On day 8, splenocytes were harvested from vaccinated mice, and CD4⁺ T cells were purified and analyzed for a Vβ11 (y-axis), b CD44 (y-axis), and a, b FOXP3 expression (x-axis). To assess generality of these findings (c, d), data from seven controlled experiments (analysis of PBMC ranging from day 4 to day 7) were pooled to assess 2D2-FIG mice vaccinated SC with saline (n = 24), 4 nmol of GMCSF-MOG (n = 24), or 4 nmol of GMCSF-NFM (n = 25) for total Treg percentages (c) and CD44⁺ Treg percentages (d) among CD4⁺ T cells. a, b, e, f Purified 2D2-FIG splenic T cells (25,000/well) from each vaccinated mouse were cultured in duplicate with 200,000 irradiated naïve splenocytes (C57BL/6) and designated concentrations (x-axis) of e MOG^35–55 and f NFM^13–37. Cultures were pulsed with 1 μCi of [³H]thymidine during the last 24 h of a 3-day culture. These data are representative of two independent experiments. Statistical significance was analyzed by use of a one-way ANOVA. (*p < 0.05, **p < 0.01, ***p < 0.001). Error bars represent standard deviations