Figure 6

UM-3003 enhances DTaP specific IgG2c production and Th1/Th17 polarization in neonatal mice. (A) 7 days old C57BL/6 were vaccinated (prime/ boost) with DTaP (1/100th of the human dose) ± UM-3003 or UM-3003 at 10 µg per mouse in alum absorbed formulations. Serum was harvested 14 days following boost (DOL 28). Anti-FHA (B–E) and anti-PRN (F–I) serum total IgG (B, F) titers, IgG1 (C, G) and IgG2c (D, H) were measured by ELISA. PBS control groups were represented as dotted line. (E, I) IgG2c/IgG1 ratios for FHA and PRN respectively. (J, K) Splenic CD4⁺ T cell responses after FHA and PRN stimulation. Four weeks following booster, splenocytes from vaccinates were isolated and stimulated with 2 μg/ml of either FHA or PRN along with CD28 (1 μg/ml) and CD49d (1 μg/ml) for 12 h followed by 6 h of BFA stimulation. After stimulation, cells were harvested and stained (intracellular cytokine staining) and analyzed by flow cytometry. Plots were gated on CD44⁺ CD4⁺ lymphocytes and analyzed for all combinations of simultaneous IFNγ, IL-4/5 and IL-17A productivity. (L) Fold change of FHA specific IFNγ production in splenic CD4⁺ T cell relative to DTaP are shown. Statistical comparison was performed either using one-way ANOVA (C, E) or nonparametric Kruskal–Wallis test corrected for multiple comparisons; *p < 0.033, **p < 0.002, ***p < 0.001 (n = 9–13 per group), with comparison to DTaP alone. Study is inclusive of two independent repeats.