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Antibody neutralization of microbiota-derived circulating peptidoglycan dampens inflammation and ameliorates autoimmunity

Nature Microbiology volume 4, pages 766–773 (2019)Cite this article

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Abstract

The human microbiota provides tonic signals that calibrate the host immune response1,2, but their identity is unknown. Bacterial peptidoglycan (PGN) subunits are likely candidates since they are well-known immunity-enhancing adjuvants, released by most bacteria during growth, and have been found in the blood of healthy people3,4,5,6,7. We developed a monoclonal antibody (mAb), 2E7, that targets muramyl-l-alanyl-d-isoglutamine (MDP), a conserved and minimal immunostimulatory structure of PGN. Using 2E7-based assays, we detected PGN ubiquitously in human blood at a broad range of concentrations that is relatively stable in each individual. We also detected PGN in the serum of several warm-blooded animals. However, PGN is barely detectable in the serum of germ-free mice, indicating that its origin is the host microbiota. Neutralization of circulating PGN via intraperitoneal administration of 2E7 suppressed the development of autoimmune arthritis and experimental autoimmune encephalomyelitis in mice. Arthritic NOD2−/− mice lacking the MDP sensor did not respond to 2E7, indicating that 2E7 dampens inflammation by blocking nucleotide-binding oligomerization domain-containing protein 2 (NOD2)-mediated pathways. We propose that circulating PGN acts as a natural immune potentiator that tunes the host immune response; altering its level is a promising therapeutic strategy for immune-mediated diseases.

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Fig. 1: Characterization of 2E7 and development of icELISA for PGN quantification.
Fig. 2: PGN levels in the serum.
Fig. 3: Increasing the level of circulating PGN promotes CIA development.
Fig. 4: Therapeutic effect of 2E7 on CAIA.

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Data availability

All data that support the findings of this study are either included in this published article and its Supplementary Information or available from the corresponding author upon request.

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Acknowledgements

We thank N.A.R. Gow, J. Heitman, and D. Ko for suggestions on the manuscript and helpful discussions. We thank N. Kaliaperumal and J. Connolly for their kind help with the Luminex and flow cytometry analyses, and W.E. Goldman for providing TCT. This work was supported by National Medical Research Council grant no. BMRC/BnB/0001b/2012 awarded to Y.W., L.C., and N.P.

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Author notes

  1. These authors contributed equally: Zhenxing Huang, Jianhe Wang, Xiaoli Xu

Authors and Affiliations

  1. Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore, Singapore

    Zhenxing Huang, Jianhe Wang, Xiaoli Xu, Haishan Wang, Yuan Qiao, Wern Cui Chu & Yue Wang

  2. Integrative Microbiology Research Centre, South China Agricultural University, Guangzhou, China

    Jianhe Wang

  3. Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore

    Shengli Xu & Kong-Peng Lam

  4. Division of Infectious Diseases, University Medicine Cluster, National University Health System, Singapore, Singapore

    Louis Chai

  5. Department of Haematology-Oncology, National University Cancer Institute, Singapore, Singapore

    Louis Chai

  6. Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

    Louis Chai

  7. Singapore Immunology Network, Agency for Science, Technology and Research, Singapore, Singapore

    Fabien Cottier & Norman Pavelka

  8. Department of Internal Medicine, Radboud University Medical Centre, Nijmegen, the Netherlands

    Marije Oosting, Leo A. B. Joosten & Mihai Netea

  9. Clinical Immunology Laboratory, Tan Tock Seng Hospital, Singapore, Singapore

    Carol Yee Leng Ng

  10. Department of Rheumatology, Allergy and Immunology Laboratory, Tan Tock Seng Hospital, Singapore, Singapore

    Khai Pang Leong

  11. Lee Kong Chian School of Medicine, Nanyang Technology University, Singapore, Singapore

    Parag Kundu & Sven Pettersson

  12. Singapore Centre for Environmental Life Sciences Engineering, Nanyang Technology University, Singapore, Singapore

    Parag Kundu & Sven Pettersson

  13. Department of Neurobiology, Care Sciences and Society, Karolinska Institutet, Stockholm, Sweden

    Sven Pettersson

  14. Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

    Yue Wang

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  1. Zhenxing Huang
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Contributions

Z.X.H. and Y.W. conceptualized the experiments. Z.X.H. and X.L.X. performed the experiments. X.L.X. and J.H.W. raised the monoclonal antibodies. H.S.W. performed the LC–MS analysis. Y.Q. isolated PGN subunits. W.C.C. conducted PGN assays in serum. S.X. and K-P.L analyzed adaptive immune response. L.C. and F.C. collected the blood samples from healthy volunteers. M.O., L.A.B.J., and M.N. collected the blood samples from donors of European ancestry. C.Y.L.N and K.P.L collected clinical samples. P.K. and S.P. raised the germ-free mice and provided serum samples. N.P. performed the statistical analyses. Y.W., L.C., and N.P. acquired the funding. Y.W. supervised the project. Y.W. and Z.X.H. wrote the manuscript.

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Correspondence to Yue Wang.

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Supplementary Figures 1–9, Supplementary Tables 1 and 2, and Supplementary References.

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Huang, Z., Wang, J., Xu, X. et al. Antibody neutralization of microbiota-derived circulating peptidoglycan dampens inflammation and ameliorates autoimmunity. Nat Microbiol 4, 766–773 (2019). https://doi.org/10.1038/s41564-019-0381-1

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