Fig. 4: High-quality antigen-specific immune responses induced by intranasal (I.N.) immunization of the FPB NC vaccine.
Determination of tPspA-specific plasma cells (A) and memory B cells (B). Two weeks after the final vaccination, tPspA-specific IgA-secreting plasma cells from the bone marrow and memory B cells from the spleen were evaluated via the ELISPOT assay (n = 4 biological replicates). Data are presented as mean values ± SEM. Statistical differences were analyzed using one-way ANOVA. Statistical significance: *P < 0.05; **P < 0.01; ****P < 0.0001; ns = not significant. Source data are provided as a Source Data file. Detail P values are provided in the Source Data file. C Determination of the tPspA-specific IgG avidity index. One week after the 2nd and 3rd immunizations, the tPspA-specific serum IgG serum avidity was evaluated using ELISA (n = 4 biological replicates). D Determination of tPspA-specific serum IgG2a/IgG1. Two weeks after the final immunization, the tPspA-specific serum IgG2a/IgG1 ratio was evaluated using an enzyme-linked immunosorbent assay (ELISA) (n = 9 biological replicates). Data are presented as mean values ± SEM. Statistical differences between two groups were analyzed using Student’s t-test. Statistical significance: *P < 0.05; **P < 0.01; ****P < 0.0001; ns = not significant. Source data are provided as a Source Data file. Detail P values are provided in the Source Data file. E Determination of tPspA-specific IFN-γ, IL-4, and IL-17 production. Two weeks after the final immunization, splenocytes were isolated and stimulated with 1 μg/ml of tPspA for three days. Cytokine levels were then measured using ELISA (n = 3 biological replicates). Data are presented as mean values ± SEM. Statistical differences were analyzed using one-way ANOVA. Statistical significance: *P < 0.05; **P < 0.01; ****P < 0.0001; ns = not significant. Source data are provided as a Source Data file. Detail P values are provided in the Source Data file.
