RNA sequencing analysis of anti-CD3/CD28 stimulated wild type-OT1 (WT) and Lrch1-/- -OT1 (KO) CD8+ T cells

We report the differential gene expression between anti-CD3/CD28 stimulated WT and LRCH1-deficient CD8+ T cells. Improving cytotoxicity, proliferation and infiltration ability of CD8+ T cells is critical in T cell immunotherapies against tumors. This study has identified LRCH1 as a negative regulator of LAT-mediated TCR signal transduction, as LRCH1 inhibits LAT signalosome formation and facilitates the endocytosis of LAT on the plasma membrane. LRCH1-deficient CD8+ T cells are more proliferative and effective at pathogen control and tumor elimination. Importantly, CRISPR/Cas9-mediated knockout of LRCH1 in human T cells also increases IFN-? production, cell proliferation and migration ability in vitro. These data suggest LRCH1 as a potential target to improve CD8+ T cell responses against tumor and pathogens. Overall design: Splenic naïve CD8+ T cells from WT and Lrch1 KO mice were cultured with plate-bound anti-CD3 (2µg/ml) plus anti-CD28 (2µg/ml) for 12 hours. Total RNA was isolated with Trizol and subjected to RNA sequencing using an Illumina HiSeq-X-Ten sequencer.