After three kinds of mixed anesthesia, antisedan was also administered intraperitoneally in all experiments. Isolation of lymphocytes from lung tissues Mice were euthanized by anesthesia and their lungs were perfused with 5?mL cold PBS solution through the right ventricle. pathway substantially increases the basal expression of antiviral genes via the spontaneous production of type I interferon (IFN). Using a combination of CRISPR/Cas9-mediated genome editing technology and a global lipidomics analysis, we found that the decrease in monounsaturated fatty acid caused by genetic deletion of in mice was crucial for the induction of MI-503 an antiviral response through activation of the cGAS-STING pathway. These findings demonstrate the important relationship between fatty acid biosynthesis and type I IFN responses that enhances the antiviral response. was crucial for the regulation of the T cell antiviral responses induced by the cGAS-STING signaling axis. Our data therefore indicate that changes in the MUFA metabolism in T cells during computer virus infection trigger cGAS-STING-mediated IFN-I-related immune responses. Results Influenza computer virus contamination reprograms lipid metabolism in lung CD4+ T cells To elucidate the role of IFN in cellular metabolism of T cells, we first performed a RNA-sequence analysis following treatment of Th1 cells with the antiviral protein IFN. Gene ontology and pathway analyses using the NIAID DAVID and KEGG databases19 showed a significant enrichment of several functional categories, including the fatty acid biosynthetic process (Fig.?1a). A gene set enrichment analysis (GSEA) also showed the decreased expression of gene sets including the fatty acid biosynthetic process (Fig.?1b). Importantly, IFN treatment resulted in the decreased expression of and gene had been conditionally deleted in CD4+ T cells driven by the promoter (herein referred to as ACC1?/?)20. To investigate the role of ACC1 in MI-503 T cell development, we analyzed thymus and spleen derived from ACC1?/? mice. ACC1?/? mice showed normal proportion and numbers of CD4+ and CD8+ T cells in the thymus (Supplementary Fig.?2a, upper panel), whereas the proportion and numbers of CD4+ and CD8+ T cells in the spleen was slightly reduced in ACC1?/? mice MI-503 compared to ACC1+/+ mice (Supplementary Fig.?2a, lower panel). However, we did not find significant changes in the proportion of memory phenotype CD4+ T cells and na?ve CD4+ T cells between ACC1+/+ and ACC1?/? mice (Supplementary Fig.?2b). To examine the effect of deletion around the expression of ISGs, we analyzed the global gene expression profiles of Th1 cells in ACC1?/? mice. A total of 395 genes showed a greater than twofold change, including 185 up-regulated and 210 down-regulated genes in ACC1?/? Th1 cells. Importantly, the genes concerning antiviral responses were significantly enriched in ACC1?/? Th1 cells without IFN-I treatment (Fig.?2a, and Supplementary Fig.?2c). A GSEA confirmed statistically significant enrichment of IFN-I-inducible genes in Rabbit Polyclonal to Ras-GRF1 (phospho-Ser916) ACC1?/? cells (Fig.?2b). Similarly, the dramatic up-regulation of ISGs was detected when we treated Th1 cells with TOFA, an allosteric inhibitor of ACC1 (Supplementary Fig.?2d and 2e). An ontology analysis also showed that this pharmacological inhibition of ACC1 resulted in the enrichment of the pathway related to the antiviral response (Supplementary Fig.?2?f). These results indicate that this cell-autonomous basal expression of antiviral genes was induced by both genetic deletion and pharmacological inhibition of ACC1 in CD4+ T cells. Open in a separate windows Fig. 2 The fatty acid biosynthesis pathway controls the ISGs expression in CD4+ T cells.a A scatter plot of gene expression by RNA-sequencing (and sgTh1 cells. Relative expression (normalized to and sgTh1 cells and infected with x31. Computer virus titers were decided 72?h p.i. (Th1 cells supplemented with 30?M palmitic acid, 10?M stearic acid, 30?M oleic acid (e) or 10?g/ml cholesterol (f) for 72?h. g, h Survival rate (g) or weight change (h) of PR8 infected mice was assessed 1 day after administrative transfer of control or sgTh1 cells. Relative expression (normalized to and (sg(sgTh1 cells. Consistently, the supplementation of MLE-15 cells with the culture supernatant from sgTh1 cell, which contained high levels of IFN, significantly enhanced antiviral activity against influenza as compared to control or sggroups (Fig.?4c and ?and4d).4d). We therefore suspected that either the accumulation of saturated fatty acid (SFA) or the deficiency of MUFA caused the induction of a type I IFN response following the deletion of Th1 cells by the addition of oleic acid (OA), which is a kind of MUFA (Fig.?4e). However, extrinsic SFA supplementation was not able to decrease the basal ISGs expression in sgTh1 cells (Fig.?4e). The supplementation of SFA into WT Th1 cell culture did not substantially.
