Fig. 3 : Human AITL malignant Tfh cells are addicted to mitochondrial respiration.

A Heatmap for GSEA data for 52 genes implicated in mitochondrial respiration (OXPHOS gene signature; [17]) comparing GSEA data of isolated Tfh cells from AITL patients (n = 6) versus healthy donor Tfh cells (n = 7) and versus public RNAseq data from healthy Tfh cells (Tfh_public, n = 7), central memory (Tcm, n = 6), effector memory (Tem, n = 6), naive (Tn, n = 6), regulatory (T reg, n = 12) and stem cell memory (Tscm, n = 6) T cells. The corresponding GSEA for the OXPHOS signature genes in AITL Tfh is indicated. For all genes with enrichment score >0 (black bars in the pink zone), expression is upregulated in hAITL. Kolmogrov–Smimov (KS) test. B Heatmap for GSEA data of genes implicated in mitochondrial biogenesis for the same T-cell fractions as in A. The corresponding GSEA for the mitochondrial biogenesis pathways in AITL Tfh indicated in B is shown. C Mitochondrial pathway analysis for expression data of the same T-cell populations as mentioned in A was performed using the KEGG database. Bubble representation (Bubbles size and numbers represent the sample enrichment score (SES), p values are indicated in Figure S3C). D Mitochondrial biogenesis pathway analysis for gene expression data of the same T cell populations as mentioned in A was performed using the Reactome database. Bubble representation (Bubbles size and numbers represent the sample enrichment score (SES), p values are indicated in Fig. S3D). E FACS analysis of mitochondrial content stained by mitotracker green (MTG) for healthy donor CD4+ T cells and hAITL total CD4+, CD4+ PD-1^low and CD4+ PD-1^high cells. (mean ± SD, n = 3; ***p < 0.001). F FACS analysis of ROS content stained by CellROX probe for healthy donor CD4+ T cells, hAITL total CD4+ T cells, CD4+ PD-1^low and hAITL CD4+ PD-1^high cells. (mean ± SD, n = 3; ***p < 0.001).