Figure 5.

Antigen-specific cellular responses in BALB/c mice heterologous prime-boost immunized with Ad5.SARS-CoV-2 S1N. BALB/c mice were immunized as follows: Group 1 served as control with SC AdΨ5 (1 × 10¹⁰ v.p.) prime and IN AdΨ5 (1 × 10¹⁰ v.p.) boost. Group 2 prime SC Ad.S1N (1 × 10¹⁰ v.p.) and IN Ad.S1N (1 × 10¹⁰ v.p.) boost. Group 3 prime IN Ad.S1N (1 × 10¹⁰ v.p.) and SC Ad.S1N (1 × 10¹⁰ v.p.) boost. Eight weeks after initial vaccination, 5 weeks post boost, splenocytes were isolated and stimulated with SARS-CoV-2 S1 and SARS-CoV-2 Nucleoprotein PepTivator, followed by ICS and flow cytometry to identify SARS-CoV-2 S1 and Nucleoprotein-specific T cells (see Supporting information for complete gating strategy). (A) Frequencies of SARS-CoV-2 S1 and Nucleoprotein-specific CD8+ IFN-γ⁺. (B) Frequencies of SARS-CoV-2 S1 and Nucleoprotein-specific CD4⁺ IFN-γ⁺. Results are mean ± SD. Groups were compared by one-way Welch’s ANOVA, followed by Dunnett’s T3 multiple comparisons, and significant differences are indicated by *p < 0.05. Unstimulated controls are represented by circles, Nucleoprotein stimulated samples (N peptide) are represented by squares, and S1 peptide stimulated samples (S peptide) are represented by triangles. Results are from a single animal experiment. (N = 5 mice per group). This experiment was conducted once.