Figure 6.

Enhancement of anabolic metabolism in TC Tfh cells

(A–D) GC-MS analysis was conducted on Tn and Tfh cells from TC or B6 mice from two cohorts. (A) Principal-component analysis (PC2 - PC3) of the 109 annotated metabolites. PC1 was ignored as it mainly accounted for batch effects, n = 7–9. (B) Pathway enrichment analysis of the significant metabolites (FDR-corrected p < 0.25, Student’s t test) comparing TC vs. B6 Tfh, Tfh vs. Tn TC, and Tfh vs. Tn B6 cells. The TC vs. B6 Tn cell comparison did not show significant metabolite features. TC Tfh-specific pathways that are also significantly enriched in in vitro-activated Tfh cells are shown in red font (see Figure S8). (C) Hierarchical clustering analyses of significant metabolites with stringent criteria (FDR-corrected p < 0.05, one-way ANOVA) among the four indicated groups of T cells are shown by heatmap. (D) Selected metabolites differentially enriched in TC vs. B6 Tfh cells (nominal p < 0.05) are shown by boxplots for the four groups of T cells. Means ± SEM analyzed by 1-way ANOVA with Dunnett’s T3 multiple comparisons tests. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001.

(E) Metabolic pathways shared between ex vivo Tfh cells compared to Tn cells shown in (A-D) and in vitro-polarized activated T cells (Tact) and Tfh cells.²³ The heatmaps show log2FC of selected significant metabolites, with the absolute numbers labeled in each cell and values with log2FC > 2 normalized to 2. DHA: dehydroascorbic acid; P: phosphate; PE: phosphatidylethanolamine; PC: Phosphatidylcholine; N-Formyl-GAR: 5′-Phosphoribosyl-N-formylglycinamide.