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Oxysterols direct B-cell migration through EBI2

Nature volume 475, pages 519–523 (2011)Cite this article

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Abstract

EBI2 (also called GPR183) is an orphan G-protein-coupled receptor that is highly expressed in spleen and upregulated upon Epstein–Barr-virus infection1. Recent studies indicated that this receptor controls follicular B-cell migration and T-cell-dependent antibody production2,3,4,5,6. Oxysterols elicit profound effects on immune and inflammatory responses as well as on cholesterol metabolism7,8,9. The biological effects of oxysterols have largely been credited to the activation of nuclear hormone receptors10,11. Here we isolate oxysterols from porcine spleen extracts and show that they are endogenous ligands for EBI2. The most potent ligand and activator is 7α,25-dihydroxycholesterol (OHC), with a dissociation constant of 450 pM for EBI2. In vitro, 7α,25-OHC stimulated the migration of EBI2-expressing mouse B and T cells with half-maximum effective concentration values around 500 pM, but had no effect on EBI2-deficient cells. In vivo, EBI2-deficient B cells or normal B cells desensitized by 7α,25-OHC pre-treatment showed reduced homing to follicular areas of the spleen. Blocking the synthesis of 7α,25-OHC in vivo with clotrimazole, a CYP7B1 inhibitor, reduced the content of 7α,25-OHC in the mouse spleen and promoted the migration of adoptively transferred pre-activated B cells to the T/B boundary (the boundary between the T-zone and B-zone in the spleen follicle), mimicking the phenotype of pre-activated B cells from EBI2-deficient mice. Our results show an unexpected causal link between EBI2, an orphan G-protein-coupled receptor controlling B-cell migration, and the known immunological effects of certain oxysterols, thus uncovering a previously unknown role for this class of molecules.

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Figure 1: Identification and pharmacological characterization of oxysterols as ligands for EBI2.
Figure 2: Regulation of EBI ligand synthesis in mouse spleen.
Figure 3: 7α,25-OHC stimulates B-cell and T-cell migration.

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Acknowledgements

We thank J. Palmer and J. Blevitt for microscopic image acquisition. We thank J. Zhu, R. Luna and S. Nguyen for technical assistance and J. Cowden for scientific discussions.

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  1. Ning Qin & Lars Karlsson

    Present address: Present addresses: Roche R&D Center (China) Ltd., 720 Cai Lun Road, Building 5, Zhangjiang High-Tech Park, Pudong, Shanghai 201203, China (N.Q.); Regulus Therapeutics, 3545 John Hopkins Court, San Diego, California 92121, USA (L.K.).,

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  1. Johnson & Johnson Pharmaceutical Research & Development, L.L.C., 3210 Merryfield Row, San Diego, 92121, California, USA

    Changlu Liu, Xia V. Yang, Jiejun Wu, Chester Kuei, Neelakandha S. Mani, Li Zhang, Jingxue Yu, Steven W. Sutton, Ning Qin, Homayon Banie, Lars Karlsson, Siquan Sun & Timothy W. Lovenberg

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Contributions

C.L., S.S., J.W., X.V.Y., N.S.M., N.Q., L.K. and T.W.L. designed and conceptualized the research. C.L., S.S., J.W., X.V.Y., N.S.M., C.K., J.Y., S.W.S., N.Q., L.Z. and H.B. performed experiments. C.L., S.S., J.W., X.V.Y., N.S.M., C.K., L.Z., N.Q, L.K. and T.W.L. analysed the data, prepared the figures and wrote the manuscript.

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Correspondence to Changlu Liu.

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Liu, C., Yang, X., Wu, J. et al. Oxysterols direct B-cell migration through EBI2. Nature 475, 519–523 (2011). https://doi.org/10.1038/nature10226

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