Gene expression profile at single cell level of CD8+ tumor infiltrating lymphocytes from the CT26 murine colon carcinoma model subdivided by antigen binding capacity

CD8+ tumor infiltrating lymphocytes can have a range of binding strengths for a single tumor associated antigen. The impact this has on the transcriptional profile of the diverse CD8+ tumor infiltrating lymphocyte population was investigated by single cell RNA sequencing. Overall design: Cells were isolated from the CT26 colon carcinoma tumor, CD8 T cells were enriched for by magnetic isolation, and stained in preparation for flow cytometry with the gp70423-431 loaded H-2Ld MHC tetramer (tetramer). CD8 T cells were isolated by fluorescence-activated cell sorting based on their capacity to bind tetramer, into high tetramer binders, low tetramer binders, and non-tetramer binders. These populations were then analyzed using scRNAseq.