Targeting oncogene-dependent TIM-3 induces T cell reprogramming thereby enhancing graft-versus-leukemia immunity - scRNAseq

We use a mouse model for AML treatment in mice, based on injection of WEHI-3B AML cells followed by allogeneic transplantation of bone marrow and T cells. Allogeneic donor T cells will elicit anti-leukemic GVL effect. We want to decipher the importance of TIM-3 expression on T cells, therefore we used donor BM and donor T cells that contains a deletion for TIM-3 in CD8+ T cells (Havcr2fl/fl;E8icre/+). We performed single cell RNA sequencing and compared T cells isolated from Havcr2fl/fl;E8icre/+ or Havcr2fl/fl (control). A total of two mice for each group were analyzed. Two Mice received WEHI-3B AML cells and additional allogeneic BM and T cells from Havcr2fl/fl (control). Two Mice received WEHI-3B AML cells and additional allogeneic BM and T cells from Havcr2fl/fl;E8icre/+. T cells were isolated on day 23 after BMT and cells were stained with H2K-b (donor haplotype) antibody (Biolegend, reference 116603) labelled with an oligotag (TotalSeq™-B0952, Biolegend, reference 405287), allowing scRNA-seq analysis of selected engrafted donor T cells.