Fig. 2: Booster-induced interferon signaling positively correlates with Th1 polarization.

A Volcano plots showing the log2(fold change) and −log10(FDR) of the differentially expressed genes (DEG) of day 1, 3, 7, and 14 post/pre-booster with downregulated genes in blue and upregulated genes in red (FDR <0.05). The number and percentage of DEG of the total analysis are depicted in the bar chart. B Left: correlation analysis using the DEG changes and T cell polarization (IFN-γ/IL-5 SFC). Dotplot of most significant correlate (GM2A day 1/pre-b with Th1 polarization on day 28) whereby the shaded region represents the 95% confidence interval (CI) around the regression line. P value was calculated by a two-sided Spearman correlation test without multiple testing correction. Middle: the number of significant correlates per time point comparison (P < 0.05) and its percentage of the total number of DEG of that time point. Right: a venn diagram depicting the number of correlates from the DEG changes on day 1/pre-b that enclosed a significant correlation with Th1 polarization pre vs post-booster including a list of the overlapping correlates. C The top 15 gene sets identified by Gene Ontology (GO) gene set enrichment analysis using the 294 DEG (day 1/pre-b) that also showed a positive correlation with the T cell polarization on day 28. D The top five known motifs identified by HOMER motif enrichment analysis using the 294 DEG (day 1/pre-b) that also showed a positive correlation with the T cell polarization on day 28. E STAT2 gene expression (TPM) over time with the black line indicating the mean expression (left) and correlation (right) between STAT2 (day 1/pre-b) and T cell polarization on day 28 (IFN-γ/IL-5 SFC) whereby the shaded region represents the 95% CI around the regression line of aP and wP groups combined (“All”). P value was calculated by a two-sided Spearman correlation test without multiple testing correction. F Heatmap depicting Spearman correlation coefficients (rs) of selected transcriptional changes (day 1/pre-b) vs T cell polarization pre- and post-booster. Insignificant correlates are crossed (P > 0.05). RNA n = 56 donors, correlation analysis n = 44 donors. A–F aP and wP-primed individuals were pooled for this analysis.