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Deleting interleukin-10 from myeloid cells exacerbates atherosclerosis in Apoe−/− mice

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Abstract

Atherosclerosis is initiated by subendothelial retention of lipoproteins and cholesterol, which triggers a non-resolving inflammatory process that over time leads to plaque progression in the artery wall. Myeloid cells and in particular macrophages are the primary drivers of the inflammatory response and plaque formation. Several immune cells including macrophages, T cells and B cells secrete the anti-inflammatory cytokine IL-10, known to be essential for the atherosclerosis protection. The cellular source of IL-10 in natural atherosclerosis progression is unknown. This study aimed to determine the main IL10-producing cell type in atherosclerosis. To do so, we crossed VertX mice, in which IRES-green fluorescent protein (eGFP) was placed downstream of exon 5 of the Il10 gene, with atherosclerosis-prone Apoe−/− mice. We found that myeloid cells express high levels of IL-10 in VertX Apoe−/− mice in both chow and western-diet fed mice. By single cell RNA sequencing and flow cytometry analysis, we identified resident and inflammatory macrophages in atherosclerotic plaques as the main IL-10 producers. To address whether IL-10 secreted by myeloid cells is essential for the protection, we utilized LyzMCre+Il10fl/fl mice crossed into the Apoe−/− background and confirmed that macrophages were unable to secrete IL-10. Chow and western diet-fed LyzMCre+Il10fl/fl Apoe−/− mice developed significantly larger atherosclerotic plaques as measured by en face morphometry than LyzMCre−Il10 fl/flApoe−/−. Flow cytometry and cytokine measurements suggest that the depletion of IL-10 in myeloid cells increases Th17 cells with elevated CCL2, and TNFα in blood plasma. We conclude that macrophage-derived IL-10 is critical for limiting atherosclerosis in mice.

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All data and methods used in the analysis and materials used to conduct the research will be made available to any researcher for the purpose of reproducing the results or replicating the procedures. All data, methods, and materials are available on personal request at the La Jolla Institute for Immunology, CA.

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Acknowledgements

We thank Jacqueline Miller for initial support with maintaining the mouse colony.

Funding

This work was supported by grants to M.O. from the American Heart Association grant (AHA18POST34060251), and grant (CDA 941152) and the Conrad Prebys Foundation Award. D.W. was supported by the Deutsche Forschungsgemeinschaft (WO 1994/1-1) and received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant agreement No 853425). H.W. was supported by Deutsche Forschungsgemeinschaft (SFB TRR259 (397484323) and CCRC GRK2407 (360043781 to HW)). K.L. was supported by grant P01 HL136275. The Zeiss LSM 880 confocal microscope was funded by NIH S10OD021831.

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Authors and Affiliations

  1. La Jolla Institute for Immunology, 9420 Athena Circle Drive, La Jolla, CA, 92037, USA

    Marco Orecchioni, Vasantika Suryawanshi, Jeffrey Makings & Klaus Ley

  2. Cardiology and Angiology I, Medical Center–University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany

    Dennis Wolf

  3. Department of Cardiology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany

    Holger Winkels

  4. Division of Vaccine Science, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Kouji Kobiyama

  5. Histology and Microscopy Core Facility, La Jolla Institute for Immunology, La Jolla, CA, 92037, USA

    William B. Kiosses

  6. Department of Bioengineering, University of California San Diego, La Jolla, CA, 92093, USA

    Klaus Ley

  7. Immunology Center of Georgia (IMMCG), Augusta University, Augusta, GA, 30912, USA

    Klaus Ley

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  1. Marco Orecchioni
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Contributions

M.O., D.W. and K.L. contributed to the study conception and design. Material preparation, data collection and analysis were performed by M.O., D.W., V.S., H.W., K.K., J.M., and W.B.K.. The first draft of the manuscript was written by M.O. and K.L. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Marco Orecchioni or Klaus Ley.

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On behalf of all authors, the corresponding author states that there is no conflict of interest.

Ethical approval

All in vivo experiments followed guidelines of the La Jolla Institute for Immunology (LJI) Animal Care and Use Committee. Approval for use of rodents was obtained from LJI according to criteria outlined in the Guide for the Care and Use of Laboratory Animals from the National Institutes of Health.

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Orecchioni, M., Wolf, D., Suryawanshi, V. et al. Deleting interleukin-10 from myeloid cells exacerbates atherosclerosis in Apoe−/− mice. Cell. Mol. Life Sci. 80, 10 (2023). https://doi.org/10.1007/s00018-022-04649-9

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