Fig. 6

The Zn-RS adjuvant formulation promoted Th1/Th2/Th9/Th17 cytokine release. Young adult (6-week-old) and aged (15-month-old) C57BL/6 mice were subcutaneously injected with 10 μg of the STFK1628x protein (formulated in the Al-001 or Zn-RS adjuvant) following either a single or double immunization. Spleen and lymph node tissues were harvested 2 weeks post-primary injection. Flow cytometric analysis of cytokine-producing T-cell subsets: A IFN-γ⁺CD4⁺ T helper (Th1) cells, IL-4⁺CD4⁺ T helper (Th2) cells, IFN-γ⁺CD8⁺ T cells, and IL-4⁺CD8⁺ T cells. B IL-9⁺CD4⁺ T helper (Th9) cells, IL-17⁺CD4⁺ T helper (Th17) cells, IL-9⁺CD8⁺ T cells, and IL-17⁺CD8⁺ T cells. The data are presented as the means ± SDs (n = 6/group). Statistical significance was tested by the Mann‒Whitney U test for pairwise comparisons and one-way ANOVA for multiple group comparisons. *p < 0.05, **p < 0.01, ***p < 0.001