BioProject

ACh was originally isolated from spleen back in 1929, however, its contribution to immune regulation has only recently been appreciated. More...

ACh was originally isolated from spleen back in 1929, however, its contribution to immune regulation has only recently been appreciated. Subsets of both T and B lymphocytes have been found to express choline acetyltransferase (Chat) and produce ACh. To date, Chat-expressing T cells have been described as relaying neural signals in the spleen to modulate immune responses; regulating blood pressure; and promoting anti-viral immune responses. In a murine multi-hit model of hepatocellular carcinoma, we observed activation of the adaptive immune response and induction of Chat-expressing CD4+ T cells. These cholinergic T cells curtail the development of liver cancer by supporting anti-tumor immune responses. We used single-cell RNA sequencing to investigate the transcriptome and TCR repertoire of the Chat-expressing CD4+ T cells in healthy and HCC-bearing livers. Overall design: HCC was induced in Chat-GFP mice by Myc overexpression and combined Trp53 and Pten ablation. HCC livers (from two male and two female mice) and control livers (from sex-matched littermates) were collected for hepatic mononuclear cell (MNC) isolation. Hepatic MNCs were stained with antibodies for CD4, CD8, CD19, TCR-β, NK1.1, CD45 and CD1d tetramer, as well as with barcode antibodies (TotalSeq-C0304, C0305, C0306, or C0307) to hashtag cells from individual mice of the HCC or control group. CD45+DAPI-NK1.1-CD1dTetramer-CD19-TCR-β+CD4+CD8-GFP+ (Chat-GFP+ CD4 T cells) and CD45+DAPI-NK1.1-CD1dTetramer-CD19-TCR-β+CD4+CD8-GFP- (Chat-GFP- CD4 T cells) populations were sorted. The two CD4 T cell compartments were individually sorted from each mouse. The same CD4 T compartments from mice under the same treatment were pooled into 4 samples: control Chat-GFP+, control Chat-GFP-, HCC Chat-GFP+ and HCC Chat-GFP-. After sorting and pooling, the samples were immediately submitted to the Princess Margaret Genomics Centre for downstream processing. Less...