A novel role for pro-coagulant microvesicles in the early host defense against streptococcus pyogenes

PLoS Pathog. 2013;9(8):e1003529. doi: 10.1371/journal.ppat.1003529. Epub 2013 Aug 1.

Abstract

Previous studies have shown that stimulation of whole blood or peripheral blood mononuclear cells with bacterial virulence factors results in the sequestration of pro-coagulant microvesicles (MVs). These particles explore their clotting activity via the extrinsic and intrinsic pathway of coagulation; however, their pathophysiological role in infectious diseases remains enigmatic. Here we describe that the interaction of pro-coagulant MVs with bacteria of the species Streptococcus pyogenes is part of the early immune response to the invading pathogen. As shown by negative staining electron microscopy and clotting assays, pro-coagulant MVs bind in the presence of plasma to the bacterial surface. Fibrinogen was identified as a linker that, through binding to the M1 protein of S. pyogenes, allows the opsonization of the bacteria by MVs. Surface plasmon resonance analysis revealed a strong interaction between pro-coagulant MVs and fibrinogen with a KD value in the nanomolar range. When performing a mass-spectrometry-based strategy to determine the protein quantity, a significant up-regulation of the fibrinogen-binding integrins CD18 and CD11b on pro-coagulant MVs was recorded. Finally we show that plasma clots induced by pro-coagulant MVs are able to prevent bacterial dissemination and possess antimicrobial activity. These findings were confirmed by in vivo experiments, as local treatment with pro-coagulant MVs dampens bacterial spreading to other organs and improved survival in an invasive streptococcal mouse model of infection. Taken together, our data implicate that pro-coagulant MVs play an important role in the early response of the innate immune system in infectious diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Coagulation / immunology*
  • CD11b Antigen / immunology*
  • CD11b Antigen / metabolism
  • CD18 Antigens / immunology*
  • CD18 Antigens / metabolism
  • Cell-Derived Microparticles / immunology*
  • Cell-Derived Microparticles / metabolism
  • Cell-Derived Microparticles / microbiology
  • Cell-Derived Microparticles / ultrastructure
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Electron, Transmission
  • Streptococcal Infections / immunology
  • Streptococcal Infections / metabolism*
  • Streptococcal Infections / pathology
  • Streptococcus pyogenes / immunology*
  • Streptococcus pyogenes / metabolism
  • Streptococcus pyogenes / ultrastructure

Substances

  • CD11b Antigen
  • CD18 Antigens
  • ITGAM protein, human

Grants and funding

This work was supported by a grant from the Deutsche Forschungsgemeinschaft (project OE 547/2-1, awarded to SO), by a grant from the Medical Faculty of the University of Rostock in the framework of the FORUN program 2012, and in part by the Swedish Research Council (project 7480) and foundations of Alfred Österlund and Crafoord, Lund (Sweden). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.