Fig. 3: Recombinant proteins induced CD4⁺ and CD8⁺ T-cell responses.

A Splenocytes isolated from immunized mice in each group on day 14 were examined by flow cytometry. The clusters of CD3⁺CD4⁺ and CD3⁺CD8⁺ cells represented the CD4⁺ (above) and CD8⁺ T cells (below). B Summary data for the fractions of CD4⁺ and CD8⁺ T cells in different groups. The data and error bars represent the means ± SDs (n = 3 biologically independent replicates). Statistical significance was calculated by one-way ANOVA with Tukey’s multiple comparison tests. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, p > 0.05. C Splenocytes were plated in ELISpot wells, and IFN-γ and IL-4 cytokine-producing cells were detected. D Frequencies of IFN-γ- and IL-4 cytokine-producing cells on day 14. The data and error bars represent the means ± SDs (n = 3 biologically independent replicates). Statistical significance was calculated by one-way ANOVA with Tukey’s multiple comparison tests. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, p > 0.05. E–I Production of cytokines, including IL-4, IL-6, IL-13, TNF-α, and IFN-γ, in the serum of patients treated with the mRNA vaccines. All the data and error bars in (E–I) represent the means ± SDs (n = 3 biologically independent replicates). Statistical significance was calculated by one-way ANOVA with Tukey’s multiple comparison tests. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, p > 0.05. J Equal numbers of CFSE^high-labeled A35R-pretreated cells and CFSE^low-labeled BSA-pretreated splenocytes were mixed and injected into immunized mice. After 12 h, the splenocytes were collected to assess antigen-specific cytolytic responses using flow cytometry. K Infected cell-specific lysis in different groups. The data and error bars represent the means ± SDs (n = 3 biologically independent replicates). Statistical significance was calculated by one-way ANOVA with Tukey’s multiple comparison tests. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, p > 0.05.