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Reciprocal coupling of coagulation and innate immunity via neutrophil serine proteases

Nature Medicine volume 16, pages 887–896 (2010)Cite this article

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Abstract

Blood neutrophils provide the first line of defense against pathogens but have also been implicated in thrombotic processes. This dual function of neutrophils could reflect an evolutionarily conserved association between blood coagulation and antimicrobial defense, although the molecular determinants and in vivo significance of this association remain unclear. Here we show that major microbicidal effectors of neutrophils, the serine proteases neutrophil elastase and cathepsin G, together with externalized nucleosomes, promote coagulation and intravascular thrombus growth in vivo. The serine proteases and extracellular nucleosomes enhance tissue factor– and factor XII–dependent coagulation in a process involving local proteolysis of the coagulation suppressor tissue factor pathway inhibitor. During systemic infection, activation of coagulation fosters compartmentalization of bacteria in liver microvessels and reduces bacterial invasion into tissue. In the absence of a pathogen challenge, neutrophil-derived serine proteases and nucleosomes can contribute to large-vessel thrombosis, the main trigger of myocardial infarction and stroke. The ability of coagulation to suppress pathogen dissemination indicates that microvessel thrombosis represents a physiological tool of host defense.

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Figure 1: Neutrophil serine proteases trigger fibrin formation.
Figure 2: TFPI degradation supports coagulation during thrombus development.
Figure 3: Effect of externalized nucleosomes on TFPI function in vitro.
Figure 4: Externalized nucleosomes promote fibrin formation in vivo. (a) Visualization of extracellular nucleosomes, neutrophils and DNA in thrombi after FeCl3-induced vessel injury (20 min) using antibody to H2A-H2B–DNA (red), myeloperoxidase (MPO)-specific antibody (green) and Hoechst 33342 (blue).
Figure 5: Effect of neutrophil serine proteases on fibrin deposition during systemic infection.
Figure 6: Effect of fibrin formation on bacterial tissue invasion.

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Acknowledgements

We acknowledge support of Deutsche Forschungsgemeinschaft and Wilhelm Sander-Stiftung to B.E. and S.M. We are grateful to W. Bode, J. Roes, Peter Lohse, Pia Lohse and A. Moseman for helpful suggestions and comments. We thank G. Längst (University of Regensburg) and K. Rippe (University of Heidelberg) for preparation of isolated nucleosomes, M. Monestier (Temple University), H. Wardemann and R. Hurwitz (Max-Planck-Institut für Infektionsbiologie) for providing H2A-H2B–DNA–specific antibody, DNA-specific antibody and human neutrophil elastase–specific antibody, K. Schledzewski (University of Heidelberg) for supplying the stabilin-2–positive antibody, L. Petersen (NovoNordisk) for donating rVIIa and D. Kirchhofer (Genentech) for providing mouse tissue factor–specific antibody. L.G. was supported by Deutsche Forschungsgemeinschaft-Forschergruppe 440 and by Rudolf-Marx-Stipendium from Gesellschaft für Thrombose-und Hämostaseforschung. K.B. and C.R. were participants of Deutsche Forschungsgemeinschaft-Graduiertenkolleg 438.

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  1. Steffen Massberg, Lenka Grahl, Marie-Luise von Bruehl and Davit Manukyan: These authors contributed equally to this work.

Authors and Affiliations

  1. Deutsches Herzzentrum, Technische Universität, Munich, Germany

    Steffen Massberg, Marie-Luise von Bruehl, Michael Lorenz, Ildiko Konrad, Elisabeth Kennerknecht & Siegmund Braun

  2. Institut für Klinische Chemie, Ludwig-Maximilians-Universität, Munich, Germany

    Lenka Grahl, Davit Manukyan, Susanne Pfeiler, Kiril Bidzhekov, Avinash B Khandagale, Katja Reges, Stefan Holdenrieder, Christoph Reinhardt & Bernd Engelmann

  3. Hämostaseologie, Ludwig-Maximilians-Universität, Munich, Germany

    Davit Manukyan & Michael Spannagl

  4. Max-Planck-Institut für Infektionsbiologie, Berlin, Germany

    Christian Goosmann & Volker Brinkmann

  5. Institut für Biochemie, Justus-Liebig-Universität, Giessen, Germany

    Klaus T Preissner

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Contributions

S.M. (together with B.E.) designed and supervised the study, analyzed data and contributed to writing the manuscript, L.G. conducted in vitro and in vivo experiments and analyzed data, M.-L.v.B. conducted in vivo experiments and analyzed data, D.M. conducted in vitro experiments and analyzed data, S.P. performed histochemical analyses, C.G. and V.B. performed morphological analyses, M.L. performed protein analyses, K.B. prepared TFPI mutants, A.B.K. performed protein analyses, I.K. and E.K. conducted part of the in vivo experiments, K.R., S.H. and S.B. performed analyses on in vitro and in vivo experiments, C.R. performed protein analyses, M.S. and K.T.P. contributed to design the study and B.E. designed the study, supervised the work, analyzed data and wrote the manuscript.

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Correspondence to Bernd Engelmann.

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Massberg, S., Grahl, L., von Bruehl, ML. et al. Reciprocal coupling of coagulation and innate immunity via neutrophil serine proteases. Nat Med 16, 887–896 (2010). https://doi.org/10.1038/nm.2184

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