Fig. 6

Cytokine responses were evaluated in immunized and control groups (3 randomly selected mice from each group at each time point) on days 38 and 180 after primary immunization. Mean cytokine responses for IFN-γ, TNF, IL-10, and IL-4 in the presence of cd-HAP antigen are shown in (A–D), respectively. To remove the backgrounds, the concentration of each cytokine in negative control (by media) was subtracted from the concentration of the same cytokine of test group stimulated by antigen and delta IFN-γ, TNF, IL-10, and IL-4 were presented in the figures. One-way ANOVA analysis showed significant mean difference for the level of IFN-γ, TNF, and IL-10 cytokines between vaccine groups 1–5. *P < 0.0001. The bars and error bars show the mean concentrations (pg/ml) of relevant cytokines and standard deviations (SD), respectively. Cytokine levels were shown on day 180 compared to day 38. E The IFN-γ/IL-4 and TNF/IL-4 ratio values, as an indicator of Th1/Th2 immune response, was calculated for each group on day 38