GSE134346
Utilizing ex vivo generated double negative T cells prevents the development and progression of nonalcoholic steatohepatitis
Methods: The DNT cells incubation with CD3/CD28 antibodies were stimulated with IL-10 (50 ng/ml) for 24 hours, then transcriptome sequencing studies were performed on DNT cells RNA. Results: The distributions of up- (107) and downregulated (114) genes (= 1.5-fold difference, padj < 0.05) compared with without IL-10-treated DNT were depicted in a volcano plot. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed that changed genes from IL-10-stimulated monocytes were involved in the immune response; inflammatory response; regulation of apoptotic and proliferation; cytolysis; cytokine-cytokine receptor interaction; and T cell receptor signaling pathway, among others. Compared with control DNT cells, IL-10-treated DNT exhibited downregulated levels of cell survival and activation genes (Bcl-xl, Cytc, Epcam, and Spi1), cell cytotoxicity genes (Nkp46, Klrg1, Slamf6, Cd107a, Cd161, and Cd226), immune responses genes (Lta, Cd127, H2-Oa, and H2-Dma), and upregulated cell exhaustion genes (Pd-1, Ctla4, and Tim3).In this study, we found IL-10 stimulated DNT cells highly expressed PI3K-AKT signaling genes, such as Jak3, Pik3ap1, Pik3cb, and Sgk1, and MAPK pathway genes, such as Stmn1, Fos, Kras, Nras, and Map3k5, which indicated that IL-10 upregulated NKG2A expression in DNT cells may through these pathways. Overall design: The DNT cells stimulated with or without IL-10 were generated by deep sequencing in duplicate, using Illumina HiSeq.