Figure 4. NK cytolytic activity and specific cellular response elicited by alum-CpG-HH2-NY vaccine.

(A) The 4-h NK assay against YAC-1 targets was determined at various effector:target ratios. Error bars represent mean + SEM. **p < 0.01, ***p < 0.001. (B) The cytolytic activity of splenocytes against NY-ESO-1⁺ B16 cells was measured using a 4 h ⁵¹Cr-release assay at various E:T ratios. Error bars represent mean + SEM. ***p < 0.001. (C) The representative graphs of spot-forming cells for IFN-γ and IL-4 at one week post-immunization in the ELISpot assay. (D) The average number of IL-4/IFN-γ secreting splenocytes was calculated (n = 3, three independent experiments). Error bars represent mean + SEM. *p < 0.05, **p < 0. 01. (E) T cell analysis in splenocytes after vaccination. Intracellular staining of IFN-γ in CD4⁺and CD8⁺T cells was analyzed by FACS.