Dengue-specific CD8+ responses by AIM. Kinetics of CD8+ responses were evaluated by AIM assay in groups vehicle-GNP (n = 6), LD PepGNP-Dengue (n = 10), and HD PepGNP-Dengue (n = 10). Percentage of antigen-specific CD8+ over total CD8+, stimulated minus unstimulated specific CD8+, defined as expressing activation co-markers CD107a+CD25+ (a), CD137+CD69+ (b) or at least one co-marker (c) upon stimulation with DENV peptides (left panels) or PepGNP-Dengue (right panels). Intra-group comparison with day 0 used Friedman tests, and inter-group comparison used Kruskal–Wallis tests. p values <0.05 are indicated. Bars indicate medians and 95% confidence intervals. Number of responders indicated above the x-axis were defined as a volunteer with a positive delta (post- minus pre-vaccination) response and frequency of dengue-specific CD8+ T cell above the mean +2 SD of the AIM+CD8+ T cell response in all volunteers at day 0 (grey zone). LD = low dose (○); HD = high dose (▵); GNP = gold nanoparticles; AIM = activation-induced markers; SD = standard deviation.