Figure 4.

UM-3003 enhances DTaP specific IgG2c production and Th1 polarization in adult mice. Adult C57BL/6 mice were immunized twice, 14 days apart (A) with DTaP (1/100th of the human dose) ± UM-3003 or alum at 10 µg per mouse in different formulations (aqueous or pre-adsorbed to alum). Serum was harvested 14 days following boost (B–I). Anti-FHA (B-E) and anti-PRN (F–I) serum Ab IgG (B, F), IgG1 (C, G) and IgG2c (D, H) titers were measured by ELISA. PBS control groups were represented as dotted line. (E, I) IgG2c/IgG1 ratios for FHA and PRN respectively. (J, K) Murine CD4⁺ T cell responses after FHA and PRN stimulation. Four weeks following booster, splenocytes from DTaP and adjuvanted vaccinates were isolated, 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 to block the extracellular cytokine secretion. After stimulation, cells were harvested, 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. Statistical analysis was performed by nonparametric Kruskal–Wallis test corrected for multiple comparisons; *p < 0.033, **p < 0.002, ***p < 0.001 (n = 8–15 per group). Study is inclusive of three independent repeats.