Figure 5. Non-toxin adjuvants differentially influence the induction of antigen-specific lymphocyte cytokine responses after nasal immunization with rPA.

Mice were immunized as described in Figure 1. Splenocytes were harvested on day 42 and re-stimulated in culture at 1.25 × 10⁶ cells per well (48 well plate) with or without rPA (5 μg/ml) to induce recall cytokine secretion by antigen-specific T cells. Supernatants were collected after 5 days of re-stimulation and assayed using Bio-Plex to identify protein levels of (A) IL-4, (B) IL-5, (C) IL-17 and (D) IFNγ. This data represents one experiment for each mouse strain (n = 5 per group). Each bar represents mean pg/ml ± standard deviation. Statistical analysis was performed using log₁₀ transformed cytokine concentrations in the rPA stimulated cultures followed by ANOVA and Tukey’s multiple comparison test. The numbers above the error bars indicate which groups (1-9) are significantly different from the indicated group.