Figure 4.

The first two vaccine inoculations elicit in HD_S a TNFα/IL-2 skewed T_H profile that is attenuated by the third dose

(A) Multiparametric UMAP representation of S-specific cytokine⁺ CD4⁺ T cells based on TNFα, CD107a, IL-10, IFNγ, IL-2, and IL-17A expression at each time point, aggregated data for the HD_S and CI cohorts. The colors identify eight populations clustered by unsupervised analysis using Phenograph.

(B) Each cluster is labeled on the global UMAP.

(C) Heatmap summarizing for each cluster the MFI of each loaded parameter.

(D) Pie charts depicting the representation of each identified cluster within total cytokine⁺ CD4⁺ T cells.

(E) Longitudinal frequencies of selected cytokine⁺ CD4⁺ T cell clusters 1, 2, 4, 6, and 8 in HD_S (orange) and CI (blue) participants. Lines connect data from the same donor. The bold lines represent the median values of each cohort. Wilcoxon tests for each pairwise comparisons are shown below the graphs.

(F) Proportions of cytokine⁺ clusters 1, 2, 4, 6, and 8 among cytokine⁺ CD4⁺ T cells in HD_S and CI at D1, D2, and D3. Bars represent median ± interquartile range. Mann-Whitey tests are shown.

(G) Cohort comparison of univariate analyses. Bars represent median ± interquartile range. Statistical comparisons using a linear mixed model are shown. In (A)–(G) n = 20 HD_S, n = 26 CI participants.