Fig. 3: Immunogenicity of accelerated prime-and-trap immunization regimens in BALB/cJ mice.

a Immunization schedule. 5ug 5-day (green data) or 5ug 14-day (red data) or 1ug 14-day regimen of repRNA-PyCS (light blue data) prime followed by trap dose of 25,000 RAS, were tested in mice. Control cohorts are prime-boost repRNA-PyCS (orange data) or trap cohort (repRNA-PfCS +RAS, dark blue data). b Liver and serum from seven mice per cohort were harvested 5 h post-trap injection. Total IgG titer was analyzed by ELISA, while liver parasite burdens were analyzed by qPCR quantification. Liver-burden qPCR was evaluated using one-way ANOVA followed by Kruskal-Wallis test and Dunn’s multiple comparisons test (*p < 0.05, **p < 0.005). A lower ΔCT represents a higher parasite burden. Pearson correlation comparing the IgG titer and the liver burden for the four vaccine groups. For visualization purposes, results from all cohorts are displayed in the same graph, but each correlation coefficient was computed individually for each cohort. c Final-bleed sera were collected at endpoint, 4 weeks post-trap (day 42), to evaluate total IgG titer and IgG1, IgG2a, IgG3 subclasses for each cohort by ELISA. Ratio of IgG2a/IgG1 is indicated in bar graph. All others statistical analyses were performed using a Mann–Whitney test. The n value represents total number of mice tested per cohort, and the error bars represent SD of the mean in two independent assays. Each data point represents an individual mouse, and the bar represents the group mean.