Abstract
Epstein–Barr virus-induced gene 2 (EBI2, also known as GPR183) is a G-protein-coupled receptor that is required for humoral immune responses; polymorphisms in the receptor have been associated with inflammatory autoimmune diseases1,2,3. The natural ligand for EBI2 has been unknown. Here we describe the identification of 7α,25-dihydroxycholesterol (also called 7α,25-OHC or 5-cholesten-3β,7α,25-triol) as a potent and selective agonist of EBI2. Functional activation of human EBI2 by 7α,25-OHC and closely related oxysterols was verified by monitoring second messenger readouts and saturable, high-affinity radioligand binding. Furthermore, we find that 7α,25-OHC and closely related oxysterols act as chemoattractants for immune cells expressing EBI2 by directing cell migration in vitro and in vivo. A critical enzyme required for the generation of 7α,25-OHC is cholesterol 25-hydroxylase (CH25H)4. Similar to EBI2 receptor knockout mice, mice deficient in CH25H fail to position activated B cells within the spleen to the outer follicle and mount a reduced plasma cell response after an immune challenge. This demonstrates that CH25H generates EBI2 biological activity in vivo and indicates that the EBI2–oxysterol signalling pathway has an important role in the adaptive immune response.
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Pereira, J. P., Kelly, L. M., Xu, Y. & Cyster, J. G. EBI2 mediates B cell segregation between the outer and centre follicle. Nature 460, 1122–1126 (2009)
Gatto, D., Paus, D., Basten, A., Mackay, C. R. & Brink, R. Guidance of B cells by the orphan G protein-coupled receptor EBI2 shapes humoral immune responses. Immunity 31, 259–269 (2009)
Heinig, M. et al. A trans-acting locus regulates an anti-viral expression network and type 1 diabetes risk. Nature 467, 460–464 (2010)
Russell, D. W. The enzymes, regulation, and genetics of bile acid synthesis. Annu. Rev. Biochem. 72, 137–174 (2003)
Lathe, R. Steroid and sterol 7-hydroxylation: ancient pathways. Steroids 67, 967–977 (2002)
Russell, D. W. Oxysterol biosynthetic enzymes. Biochim. Biophys. Acta 1529, 126–135 (2000)
Bensinger, S. J. et al. LXR signaling couples sterol metabolism to proliferation in the acquired immune response. Cell 134, 97–111 (2008)
Gauwerky, C. E., Hoxie, J., Nowell, P. C. & Croce, C. M. Pre-B-cell leukemia with a t(8; 14) and a t(14; 18) translocation is preceded by follicular lymphoma. Oncogene 2, 431–435 (1988)
Reed, J. C. & Tanaka, S. Somatic point mutations in the translocated bcl-2 genes of non-Hodgkin’s lymphomas and lymphocytic leukemias: implications for mechanisms of tumor progression. Leuk. Lymphoma 10, 157–163 (1993)
Novershtern, N. et al. Densely interconnected transcriptional circuits control cell states in human hematopoiesis. Cell 144, 296–309 (2011)
Rubic, T. et al. Triggering the succinate receptor GPR91 on dendritic cells enhances immunity. Nature Immunol. 9, 1261–1269 (2008)
Bauman, D. R. et al. 25-Hydroxycholesterol secreted by macrophages in response to Toll-like receptor activation suppresses immunoglobulin A production. Proc. Natl Acad. Sci. USA 106, 16764–16769 (2009)
Diczfalusy, U. et al. Marked upregulation of cholesterol 25-hydroxylase expression by lipopolysaccharide. J. Lipid Res. 50, 2258–2264 (2009)
Honda, A. et al. Cholesterol 25-hydroxylation activity of CYP3A. J. Lipid Res. 10.1194/jlr.M014084 (2011)
Li, X. et al. Biosynthesis of the regulatory oxysterol, 5-cholesten-3β,25-diol 3-sulfate, in hepatocytes. J. Lipid Res. 48, 2587–2596 (2007)
Lund, E., Bjorkhem, I., Furster, C. & Wikvall, K. 24-, 25- and 27-hydroxylation of cholesterol by a purified preparation of 27-hydroxylase from pig liver. Biochim. Biophys. Acta 1166, 177–182 (1993)
Dulos, J., van der Vleuten, M. A., Kavelaars, A., Heijnen, C. J. & Boots, A. M. CYP7B expression and activity in fibroblast-like synoviocytes from patients with rheumatoid arthritis: regulation by proinflammatory cytokines. Arthritis Rheum. 52, 770–778 (2005)
Wang, J. H. & Tuohimaa, P. Regulation of cholesterol 25-hydroxylase expression by vitamin D3 metabolites in human prostate stromal cells. Biochem. Biophys. Res. Commun. 345, 720–725 (2006)
Park, K. & Scott, A. L. Cholesterol 25-hydroxylase production by dendritic cells and macrophages is regulated by type I interferons. J. Leukoc. Biol. 88, 1081–1087 (2010)
Liu, C. et al. Oxysterols direct B-cell migration through EBI2. Nature 10.1038/10226 (2011)
Li, D. & Spencer, T. A. Synthesis of 7α-hydroxy derivatives of regulatory oxysterols. Steroids 65, 529–535 (2000)
Acknowledgements
We would like to thank the following scientists for their support: F. Su and J.-L. Vincent (Bruxelles Erasme Hospital) for access to tissue samples from the peritonitis sheep sepsis model; D. W. Russell (University of Texas Southwestern) for access to the Ch25h−/− mouse strain; J. Carballido and T. Rubic (all Novartis) for providing an EBV-infected B-cell line and help in preparation of mouse bone-marrow-derived dendritic cells; J. C. Reed (Burnham Institute for Medical Research) for the RS11846 cells; A.-G. Letombe (Euroscreen), A. Feige and N. Beluch for technical assistance; and J. Hamon, C. Bauer, A. Glaenzel, P. Bross, V. Techer-Etienne (all Novartis) and RC Tritec Ltd (Teufen, Switzerland) for support in the development of a radioligand binding assay.
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S.H., S.N., M.-O.R., M.D. and K.S. initiated the project and developed strategy. S.H., S.N. and M.O.R. purified activity from sheep tissue extracts. W.S., D.-I.K., C.Y.C. and P.G.S. carried out corresponding work on the pig tissue. J.Z., Ri.K., I.C., Ra.K., R.F. and E.C.P. conducted mass-spectrometry and NMR experiments. S.L., M.V., I.P., D.G., D.N., W.L., A.K., Y.A.C., B.U.B., K.S. and A.W.S. designed and conducted pharmacological characterization of the receptor and bioactivity assays. S.R., C.M., F.G., Ca.S., W.S., C.Y.C., S.H. and A.W.S. proposed and/or synthesized various oxysterol ligands. T.S., Ch.S. and B.W. supported in vivo experiments with mouse models. T.Y., J.P.P., L.M.K. and J.G.C. developed the bioassay, conducted the experiments in CH25H-deficient and transduced mice and performed the B-cell chemotaxis and immunization studies. A.W.S. wrote the first draft of the manuscript, which was completed by S.H., J.Z., D.N., B.W., S.R., J.G.C., C.Y.C., K.S. and commented on by all authors.
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All authors, except for T.Y., J.P.P., L.M.K., D.I.K. and J.G.C. are current or previous employees of either Euroscreen or Novartis and some of them do hold stock or stock options in their respective companies.
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Hannedouche, S., Zhang, J., Yi, T. et al. Oxysterols direct immune cell migration via EBI2. Nature 475, 524–527 (2011). https://doi.org/10.1038/nature10280
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DOI: https://doi.org/10.1038/nature10280
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