Complete T cell antigen suppresses CD8 ⁺ TIL exhaustion and promotes infiltration of effector memory CD8 ⁺ T cells. The MC38 tumor-bearing mice received Bpmel-SIY-OVAII cell therapy on day 6 after MC38 inoculation, and CD8⁺ TILs were subjected to scRNAseq on day 13. (A) In the upper panel, uniform manifold approximation and projection (UMAP) of CD8⁺ TILs treated with or without Bpmel-SIY-OVAII cell adjuvant (control TIL: n = 10,357 and SIY-OVAII TIL: n = 10,357). The name of each cluster and annotations of the cell types labelled with ProjecTILs were shown in the lower panel. (B) Percentages of each CD8⁺ TIL population based on the annotation by ProjecTILs. (C) A volcano plot showing differential expression of genes (DEGs) in total exhausted CD8⁺ TILs (Tex + Tpex) between the control and SIY-OVAII groups. (D) The GSEA enriched the pathway of “GOBP ALPHA BETA T CELL ACTIVATION” in CD8⁺ exhausted T cells between the control and SIY-OVAII TIL groups. (E) UMAP of total Tex and Tpex CD8⁺ TILs from the control and SIY-OVAII groups. (F) A volcano plot showing DEGs of CD8⁺ effector memory TILs between the control and SIY-OVAII groups. (G) UMAP of CD8⁺ effector memory TILs from the control or SIY-OVAII groups. (H) A volcano plot showing DEGs between clusters 4 vs. 2 plus 3. (I) UAMP dimension reduction was performed using Monocle3 with Seurat cluster labels (left), and pseudo-time values were calculated and plotted (right). GSEA, gene set enrichment analysis.