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Mrp8 and Mrp14 are endogenous activators of Toll-like receptor 4, promoting lethal, endotoxin-induced shock

Nature Medicine volume 13, pages 1042–1049 (2007)Cite this article

Abstract

To identify new components that regulate the inflammatory cascade during sepsis, we characterized the functions of myeloid-related protein-8 (Mrp8, S100A8) and myeloid-related protein-14 (Mrp14, S100A9), two abundant cytoplasmic proteins of phagocytes. We now demonstrate that mice lacking Mrp8-Mrp14 complexes are protected from endotoxin-induced lethal shock and Escherichia coli–induced abdominal sepsis. Both proteins are released during activation of phagocytes, and Mrp8-Mrp14 complexes amplify the endotoxin-triggered inflammatory responses of phagocytes. Mrp8 is the active component that induces intracellular translocation of myeloid differentiation primary response protein 88 and activation of interleukin-1 receptor–associated kinase-1 and nuclear factor-κB, resulting in elevated expression of tumor necrosis factor-α (TNF-α). Using phagocytes expressing a nonfunctional Toll-like receptor 4 (TLR4), HEK293 cells transfected with TLR4, CD14 and MD2, and by surface plasmon resonance studies in vitro, we demonstrate that Mrp8 specifically interacts with the TLR4-MD2 complex, thus representing an endogenous ligand of TLR4. Therefore Mrp8-Mrp14 complexes are new inflammatory components that amplify phagocyte activation during sepsis upstream of TNFα–dependent effects.

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Figure 1: Effects of LPS on Mrp14−/− mouse bone marrow cells (BMCs).
Figure 2: Extracellular Mrp8-Mrp14 and Mrp8 augment LPS signaling.
Figure 3: Mrp8 and LPS signaling are dependent on MAP kinase p38, ERK and PKC.
Figure 4: Mrp8 induces TLR4–dependent TNF-α expression.
Figure 5: Binding of Mrp8 to the TLR4 signaling complex.
Figure 6: Mrp8-Mrp14 and Mrp8 promote LPS-induced shock.

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Acknowledgements

We thank C. Sachs, H. Berheide, E. Nattkemper, J. Daalhuisen and K. Övermöhle for excellent technical assistance; P. Björk (Active Biotech, Lund) for performing surface plasmon resonance studies and W. Falk (Department of Internal Medicine I, University of Regensburg, Germany) for providing D. melanogaster Schneider 2 (S2) cells expressing mouseTLR4-MD2. This work was supported by the Interdisciplinary Clinical Research Centre of the University of Muenster (grant Ro20/12/06 to J.R.) and the German Research Foundation (grant SFB293 A16 to J.R.).

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Authors and Affiliations

  1. Institute of Experimental Dermatology, University of Münster, Münster, D-48129, Germany

    Thomas Vogl, Nadja Leukert, Wolfgang Nacken, Clemens Sorg & Johannes Roth

  2. Interdisciplinary Center of Clinical Research, University of Münster, Münster, D-48129, Germany

    Klaus Tenbrock, Dirk Foell & Johannes Roth

  3. Department of Pediatrics, University of Münster, D-48129 Münster, Germany.,

    Klaus Tenbrock, Dirk Foell & Johannes Roth

  4. Institute of Molecular Virology, University of Münster, Münster, D-48129, Germany

    Stephan Ludwig & Christina Ehrhardt

  5. Center of Infection and Immunity, Academic Medical Center, University of Amsterdam, Amsterdam, 1105, AZ, The Netherlands

    Marieke A D van Zoelen & Tom van der Poll

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Contributions

T.V. designed the study and the experiments, performed animal studies and experiments and wrote the manuscript. K.T. performed the ChIP experiments and analyzed data. S.L. and C.E. performed cell-culture experiments and transfection studies. N.L. and D.F. performed the immunohistological analyses and flow cytometry. M.A.D.v.Z. and T.v.d.P. performed and analyzed animal studies. W.N. generated the Mrp14−/− mice and assisted with the animal studies. C.S. assisted with the design of experiments. J.R. designed the study and experiments, analyzed data, supervised the study and wrote the manuscript.

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Correspondence to Johannes Roth.

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Vogl, T., Tenbrock, K., Ludwig, S. et al. Mrp8 and Mrp14 are endogenous activators of Toll-like receptor 4, promoting lethal, endotoxin-induced shock. Nat Med 13, 1042–1049 (2007). https://doi.org/10.1038/nm1638

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