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Interleukin 18 function in atherosclerosis is mediated by the interleukin 18 receptor and the Na-Cl co-transporter

Nature Medicine volume 21, pages 820–826 (2015)Cite this article

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Abstract

Interleukin-18 (IL18) participates in atherogenesis through several putative mechanisms1,2. Interruption of IL18 action reduces atherosclerosis in mice3,4. Here, we show that absence of the IL18 receptor (IL18r) does not affect atherosclerosis in apolipoprotein E–deficient (Apoe−/−) mice, nor does it affect IL18 cell surface binding to or signaling in endothelial cells. As identified initially by co-immunoprecipitation with IL18, we found that IL18 interacts with the Na-Cl co-transporter (NCC; also known as SLC12A3), a 12-transmembrane-domain ion transporter protein preferentially expressed in the kidney5. NCC is expressed in atherosclerotic lesions, where it colocalizes with IL18r. In Apoe−/− mice, combined deficiency of IL18r and NCC, but not single deficiency of either protein, protects mice from atherosclerosis. Peritoneal macrophages from Apoe−/− mice or from Apoe−/− mice lacking IL18r or NCC show IL18 binding and induction of cell signaling and cytokine and chemokine expression, but macrophages from Apoe−/− mice with combined deficiency of IL18r and NCC have a blunted response. An interaction between NCC and IL18r on macrophages was detected by co-immunoprecipitation. IL18 binds to the cell surface of NCC-transfected COS-7 cells, which do not express IL18r, and induces cell signaling and cytokine expression. This study identifies NCC as an IL18-binding protein that collaborates with IL18r in cell signaling, inflammatory molecule expression, and experimental atherogenesis.

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Figure 1: Identification of alternative IL18-binding proteins.
Figure 2: NCC expression and characterization.
Figure 3: IL18r and NCC function in atherosclerosis.
Figure 4: NCC mediates IL18 signaling and downstream cytokine and chemokine production in macrophages, COS-7 cells and FlpIn-293 cells.

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Acknowledgements

This study is supported by grants from the US National Heart, Lung, and Blood Institute (HL60942, HL81090, HL88547 to G.-P.S.; HL34636, HL80472 to P.L.) and by an American Heart Association Established Investigator Award (0840118N to G.-P.S.).

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Author notes

  1. Jing Wang and Chongxiu Sun: These authors contributed equally to this study.

Authors and Affiliations

  1. Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA

    Jing Wang, Chongxiu Sun, Norbert Gerdes, Conglin Liu, Mengyang Liao, Jian Liu, Michael A Shi, Aina He, Yi Zhou, Galina K Sukhova, Huimei Chen, Jie Zhang, Xiang Cheng, Qiang Ke, Peter Libby & Guo-Ping Shi

  2. Department of Pathophysiology, State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Tsinghua University, Beijing, China

    Jing Wang

  3. Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, China

    Chongxiu Sun

  4. Institute for Cardiovascular Prevention (IPEK), Ludwig-Maximilians University Munich, Munich, Germany

    Norbert Gerdes

  5. Institute of Clinical Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China

    Conglin Liu

  6. Institute of Cardiology, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, China

    Mengyang Liao & Xiang Cheng

  7. Departments of Cardiology and Geriatrics, Graduate School of Medicine, Nagoya University, Nagoya, Japan

    Xian Wu Cheng, Masafumi Kuzuya & Toyoaki Murohara

  8. Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA

    Mengmeng Jiang & Gary E Shull

  9. Division of Nephrology and Hypertension, Oregon Health and Science University, Portland, Oregon, USA

    Shaunessy Rogers, Chao-Ling Yang & David H Ellison

  10. Department of Physiology, Radboud University Nijmegen Medical Centre, Nijmegen, the Netherlands

    Sabina Jelen & René Bindels

  11. Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA

    Petr Jarolim

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  1. Jing Wang
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Contributions

J.W. and C.S. performed most of the experiments. N.G. completed the original IL18 and IL18r mutant mouse analysis. C.L., M.L., M.A.S., A.H., Y.Z., H.C., J.Z., X.C. and Q.K. performed RT-PCR, lesion analysis, cell culture and plasma ELISA. J.L. helped with the NCC cDNA cloning. G.K.S. performed immunostaining. X.W.C., M.K., T.M. and P.L. helped with experimental design, writing and data interpretation. M.J. and G.E.S. provided the NCC mutant mice. S.R., C.-L.Y. and D.H.E. provided the NCC monoclonal antibody and performed the 293 cell experiments. S.J. and R.B. made the human NCC mutant constructs. P.J. measured plasma Mg and K. G.-P.S. designed and performed the experiments and wrote the manuscript.

Corresponding author

Correspondence to Guo-Ping Shi.

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Wang, J., Sun, C., Gerdes, N. et al. Interleukin 18 function in atherosclerosis is mediated by the interleukin 18 receptor and the Na-Cl co-transporter. Nat Med 21, 820–826 (2015). https://doi.org/10.1038/nm.3890

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