Fig. 5: Neonatal mice germinal center response to CpG containing PPS14-TT vaccine.
From: IL6 suppresses vaccine responses in neonates by enhancing IL2 activity on T follicular helper cells
C57BL/6J mice were immunized i.p. with PPS14-TT (PBS) or PPS14-TT + CpG (CpG) and splenocytes were analyzed by FACS at 7 dpi. a Representative dot plots from 24 h post immunization depict the FMO control and the percentages of IL-6+ cells on total splenocytes. Mean percentages of IL-6+ cells among splenocytes are plotted (n = 5). b Representative dot plots from 24 h post immunization depict the FMO control and the percentages of IL-6+ cells on CD11c+ cells. Mean percentages of CD11c+IL-6+ subsets are plotted (n = 5). c Splenocytes from 7 dpi were in vitro stimulated with PMA/Ion for 4 h and TFH cells were analyzed. Representative contour plots depict the percentages of IL-2+ cells among TFH (CXCR5hiPD-1hi) population pre-gated on CD4+FoxP3− cells. Mean percentages of IL-2+ cells among TFH cells are plotted (n = 5). d, e Splenocytes from 7 dpi were pre-gated on CD4+CXCR5hiPD-1hi TFH cells. Representative contour plots depict percentages of IL-2Rα (d) or IL-2Rβ (e) expressing FoxP3+ and FoxP3− cells on TFH cells. Mean percentages of IL-2Rα+ and IL-2Rβ+ cells among TFH cells are also plotted (n = 6). f Splenocytes from 7 dpi were stimulated with or without recombinant IL-2 for 15 min, followed by intracellular staining for p-STAT5. Representative contour plots depict the percentages of p-STAT5+ cells among FoxP3+ and FoxP3− TFH (pre-gated on CD4+CXCR5hiPD-1hi) cells. Mean percentages of p-STAT5+ cells among TFH cells are plotted (n = 5). Experiments were performed twice. Unpaired student’s t-test and One-Way ANOVA were used for all comparisons; data represented as mean ± SEM are shown. P values < 0.05 were considered statistically significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
