Fig 1: Evaluation of antigen‐specific IFN‐γ, IL‐2, and TNF‐α secretion in BALB/c mice immunized with the Ad5Spike vaccine vector. Female BALB/c mice (6–8 weeks old; n = 5 per group per timepoint) were immunized intraperitoneally with varying doses of the Ad5Spike recombinant adenoviral vector containing 108, 1010, or 101 2 viral particles at two time points: 30 days (left panels) and 90 days (right panels) post‐immunization. (A) IFN‐γ levels in mouse sera were measured using ELISA. (B) IFN‐γ–secreting splenocytes were quantified by ELISpot assay. (C) IL‐2–secreting splenocytes and (D) TNF‐α–secreting splenocytes were measured by ELISpot assays. Data are presented as mean ± SEM, with each data point representing an individual mouse. Normality was assessed using the Shapiro–Wilk test. Statistical analysis was performed using one‐way ANOVA followed by Tukey's multiple comparisons test. The AdGFP‐immunized group served as the reference control. *p < 0.05, ****p < 0.0001.