Fig. 3: Adoptive Treg cell therapy-induced Th17 cells exhibit high pathogenicity.

A UMAP plot of all Th17 cells colored by clusters. B Bubble plot showing the top 15 GO (BP) terms (rows) for C1 upregulated gene enrichment pathways. Functional enrichment analysis was performed using clusterProfiler via over-representation analysis (ORA), which employs the hypergeometric test to calculate enrichment p-values. The Benjamini-Hochberg method controlled the false discovery rate (FDR) across all tested gene sets. C Gene set enrichment analysis (GSEA) of pathogenic Th17-related genes in C1, comparing C0. NES: normalized enrichment score. NES were computed using a weighted enrichment statistic applied to signal-to-noise-ranked genes. P-value for each gene set was estimated through gene set permutation (n = 1000). The BH method controlled the FDR across all tested gene sets. D Fraction of the different clusters in the indicated groups. E Heat map shows the average expression levels of pathogenic/non-pathogenic Th17-related genes (columns; Z normalized per column) in the indicated groups (rows, after batch correction). F Experimental scheme of the IBD model to verify the pathogenicity of Th17 cells induced by Tregs in vivo. G, H CD45.2⁺CD4⁺ effector T cells isolated from the IBD group, CD45.2⁺ CD4⁺IL17(eGFP)⁺ Th17 cells and CD45.2⁺CD4⁺IL17(eGFP)^- effector T cells isolated from the Treg cell therapy group were transferred into Rag1^-/- mice and then observed for the development of colitis. G Weight changes of Rag1^−/− mice post T cell transfer (n = 3 mice per group); mean ± SD, two-way multiple-range ANOVA test. H Representative histology images of colon sections. Scale bars, 500 μm. Data are representative of two independent experiments (G, H). Source data are provided as a Source Data file.