The glycosphingolipid, α-galactosylceramide (αGalCer), when presented by CD1d on antigen-presenting cells, efficiently activates invariant natural killer T (iNKT) cells. Thereby, it modulates immune responses against tumors, microbial and viral infections, and autoimmune diseases. Recently, the production of αGalCer by Bacteroidetes from the human gut microbiome was elucidated. Using hydrophilic interaction chromatography coupled to MS2, we screened murine intestinal tracts to identify and quantify αGalCers, and we investigated the αGalCer response to different dietary and physiologic conditions. In both the cecum and the colon of mice, we found 1-15 pmol of αGalCer per milligram of protein; in contrast, mice lacking microbiota (germ-free mice) and fed identical diet did not harbor αGalCer. The identified αGalCer contained a β(R)-hydroxylated hexadecanoyl chain N-linked to C18-sphinganine, which differed from what has been reported with Bacteroides fragilis Unlike β-anomeric structures, but similar to αGalCers from B. fragilis, the synthetic form of the murine αGalCer induced iNKT cell activation in vitro. Last, we observed a decrease in αGalCer production in mice exposed to conditions that alter the composition of the gut microbiota, including Western type diet, colitis, and influenza A virus infection. Collectively, this study suggests that αGalCer is produced by commensals in the mouse intestine and reveals that stressful conditions causing dysbiosis alter its synthesis. The consequences of this altered production on iNKT cell-mediated local and systemic immune responses are worthy of future studies.

Center for Mass Spectrometry and Optical Spectroscopy Mannheim University of Applied Sciences Mannheim Germany

Centre d'Infection et d'Immunité de Lille Inserm U1019 CNRS UMR 8204 University of Lille CHU Lille Institut Pasteur de Lille Lille France

Department of Cellular and Molecular Pathology German Cancer Research Center Heidelberg Germany

Division of Virus Associated Carcinogenesis German Cancer Research Center Heidelberg Germany

Faculty of Biosciences University of Heidelberg Heidelberg Germany

Institute of Pharmacology University of Marburg Marburg Germany

Lipid Pathobiochemistry Group Department of Cellular and Molecular Pathology German Cancer Research Center Heidelberg Germany

Skin Barrier Research Group Department of Organic and Bioorganic Chemistry Faculty of Pharmacy in Hradec Králové Charles University Hradec Králové Czech Republic

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J Lipid Res. 2019 Nov;60(11):1805-1806. doi: 10.1194/jlr.C119000437. PubMed

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