Gene expression profile at single cell level of CD44+CD8+ from tumors and PD-1+CD8+ T cells from draining LNs of untreated and aPDL-1 and CTLA-4 treated AT3-OVA-bearing mice.

Tumor-infiltrating CD8+ (TILs) exhibit heterogeneity, encompassing precursors of exhausted T (TPEX) cells and terminally exhausted T cells. We employed single cell RNA sequencing to analyze the diversity of TILs and explore the developmental and functional relationship between TPEX cells within the tumor and in those in tumor-draining lymph nodes. Lymphocytes were isolated from AT3-OVA tumors and tumor-draining lymph nodes that were either left untreated or treated 3x with aPDL-1 and aCTLA4, and single-cell suspensions were prepared. CD44+ T cells from tumors or CD44+PD-1+ CD8+ T cells from lymph nodes were sorted using Fluorescence-Activated Cell Sorting (FACS) and analyzed through single-cell RNA sequencing (scRNAseq)