Fig. 5: Distinct transcriptional profiles of PD-1⁺ alloreactive CD8⁺ T cell subsets in acute GvHD.

ScRNA-seq was performed using PD-1⁺ alloreactive CD8⁺ T cells isolated from the spleen of mice with acute GvHD at 33–35 dpt. A, B UMAP showing unsupervised Seurat clusters (A) and supervised cluster groupings based on the phenotype (B) of PD-1⁺ CD8⁺ T cells across fourteen integrated samples. C Violin plots of representative genes highly expressed in all three PD-1⁺ CD8⁺ T cell subsets or differentially expressed in each subset. D Gene set enrichment analysis for comparing specific gene signatures of PD-1⁺ CD8⁺ T cell subsets in acute GvHD to those of exhausted CD8⁺ T cell subsets in chronically infected mice (available in PRJNA497086). The depth of the color represents the normalized enrichment score. The area of the circle in the graph reflects the adjusted P-value determined by one-tailed permutation test. E Comparison of differentially expressed genes between progenitor and terminally differentiated PD-1⁺ CD8⁺ T cell subsets. Volcano plot shows fold-change (log₂) versus adjusted P-value (-log₁₀) for each gene. Significance was determined as |log₂fold-change > log₂(1.25)| and adjusted P-value < 0.01, two-sided Wilcoxon test. F KEGG pathway analysis of the highly expressed genes in each subset. A false discovery rate (FDR) < 0.05 indicates significant change. Data are representative of 10 biologically independent pooled samples. Source data are provided as a Source Data file.