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.
Vyšlo v časopise: A Novel Role for Pro-Coagulant Microvesicles in the Early Host Defense against. PLoS Pathog 9(8): e32767. doi:10.1371/journal.ppat.1003529 Kategorie: Research Article prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003529
1. EsmonCT, XuJ, LupuF (2011) Innate immunity and coagulation. J Thromb Haemost 9 Suppl 1 : 182–188 doi:10.1111/j.1538-7836.2011.04323.x
2. HaningtonPC, ZhangS-M (2011) The primary role of fibrinogen-related proteins in invertebrates is defense, not coagulation. J Innate Immun 3 : 17–27 doi:10.1159/000321882
3. KremMM, Di CeraE (2002) Evolution of enzyme cascades from embryonic development to blood coagulation. Trends Biochem Sci 27 : 67–74.
4. LoofTG, SchmidtO, HerwaldH, TheopoldU (2011) Coagulation Systems of Invertebrates and Vertebrates and Their Roles in Innate Immunity: The Same Side of Two Coins? J Innate Immun 3 : 34–40.
5. FurieB, FurieBC (1988) The molecular basis of blood coagulation. Cell 53 : 505–518.
6. ColeJN, BarnettTC, NizetV, WalkerMJ (2011) Molecular insight into invasive group A streptococcal disease. Nat Rev Micro 9 : 724–736.
7. BryantAE, Hayes-SchroerSM, StevensDL (2003) M type 1 and 3 group A streptococci stimulate tissue factor-mediated procoagulant activity in human monocytes and endothelial cells. Infect Immun 71 : 1903–1910.
8. PåhlmanLI, MalmströmE, MörgelinM, HerwaldH (2007) M protein from Streptococcus pyogenes induces tissue factor expression and pro-coagulant activity in human monocytes. Microbiology (Reading, Engl) 153 : 2458–2464.
9. Ben NasrAB, HerwaldH, Müller-EsterlW, BjörckL (1995) Human kininogens interact with M protein, a bacterial surface protein and virulence determinant. Biochem J 305 ((Pt 1)) 173–180.
10. OehmckeS, ShannonO, MörgelinM, HerwaldH (2010) Streptococcal M proteins and their role as virulence determinants. Clin Chim Acta 411 : 1172–1180.
11. HerwaldH, CramerH, MörgelinM, RussellW, SollenbergU, et al. (2004) M protein, a classical bacterial virulence determinant, forms complexes with fibrinogen that induce vascular leakage. Cell 116 : 367–379.
12. PåhlmanLI, MörgelinM, EckertJ, JohanssonL, RussellW, et al. (2006) Streptococcal M protein: a multipotent and powerful inducer of inflammation. J Immunol 177 : 1221–1228.
13. OehmckeS, MörgelinM, MalmströmJ, LinderA, ChewM, et al. (2012) Stimulation of blood mononuclear cells with bacterial virulence factors leads to the release of pro-coagulant and pro-inflammatory microparticles. Cell Microbiol 14 : 107–119.
14. GyörgyB, SzabóTG, PásztóiM, PálZ, MisjákP, et al. (2011) Membrane vesicles, current state-of-the-art: emerging role of extracellular vesicles. Cell Mol Life Sci 68 : 2667–2688.
15. BurnierL, FontanaP, KwakBR, Angelillo-ScherrerA (2009) Cell-derived microparticles in haemostasis and vascular medicine. Thromb Haemost 101 : 439–451.
16. TimárCI, LorinczAM, Csépányi-KömiR, Vályi-NagyA, NagyG, et al. (2013) Antibacterial effect of microvesicles released from human neutrophilic granulocytes. Blood 121 : 510–518.
17. ÅkessonP, SchmidtKH, CooneyJ, BjörckL (1994) M1 protein and protein H: IgGFc - and albumin-binding streptococcal surface proteins encoded by adjacent genes. Biochem J 300 ((Pt 3)) 877–886.
18. CollinM, OlsénA (2000) Generation of a mature streptococcal cysteine proteinase is dependent on cell wall-anchored M1 protein. Mol Microbiol 36 : 1306–1318.
19. MassbergS, GrahlL, Bruehl VonM-L, ManukyanD, PfeilerS, et al. (2010) Reciprocal coupling of coagulation and innate immunity via neutrophil serine proteases. Nat Med 16 : 887–896.
20. LoofTG, MörgelinM, JohanssonL, OehmckeS, OlinAI, et al. (2011) Coagulation, an ancestral serine protease cascade, exerts a novel function in early immune defense. Blood 118 : 2589–2598.
21. PlowEF, PierschbacherMD, RuoslahtiE, MarguerieG, GinsbergMH (1987) Arginyl-glycyl-aspartic acid sequences and fibrinogen binding to platelets. Blood 70 : 110–115.
22. ReidVL, WebsterNR (2012) Role of microparticles in sepsis. Br J Anaesth 109 : 503–513.
23. CimminoG, GolinoP, BadimonJJ (2010) Pathophysiological role of blood-borne tissue factor: should the old paradigm be revisited? Intern Emerg Med 6 : 29–34.
24. MorelO, MorelN, JeselL, FreyssinetJ-M, TotiF (2011) Microparticles: a critical component in the nexus between inflammation, immunity, and thrombosis. Semin Immunopathol 33 : 469–486.
25. NieuwlandR, BerckmansRJ, McGregorS, BöingAN, RomijnFP, et al. (2000) Cellular origin and procoagulant properties of microparticles in meningococcal sepsis. Blood 95 : 930–935.
26. SunH (2011) Exploration of the host haemostatic system by group A streptococcus: implications in searching for novel antimicrobial therapies. J Thromb Haemost 9 Suppl 1 : 189–194.
27. SixtM, HallmannR, WendlerO, Scharffetter-KochanekK, SorokinLM (2001) Cell adhesion and migration properties of beta 2-integrin negative polymorphonuclear granulocytes on defined extracellular matrix molecules. Relevance for leukocyte extravasation. J Biol Chem 276 : 18878–18887.
28. KanseSM, MatzRL, PreissnerKT, PeterK (2004) Promotion of leukocyte adhesion by a novel interaction between vitronectin and the beta2 integrin Mac-1 (alphaMbeta2, CD11b/CD18). Arterioscl Throm Vas 24 : 2251–2256.
29. LishkoVK, YakubenkoVP, UgarovaTP (2003) The interplay between integrins alphaMbeta2 and alpha5beta1 during cell migration to fibronectin. Exp Cell Res 283 : 116–126.
30. TalaySR (2005) Gram-positive adhesins. Contrib Microbiol 12 : 90–113.
31. KasettyG, PapareddyP, KalleM, RydengårdV, WalseB, et al. (2011) The C-terminal sequence of several human serine proteases encodes host defense functions. J Innate Immun 3 : 471–482.
32. PapareddyP, RydengårdV, PasupuletiM, WalseB, MörgelinM, et al. (2010) Proteolysis of human thrombin generates novel host defense peptides. PLoS Pathog 6: e1000857.
33. OehmckeS, ShannonO, Koeckritz-Blickwede vonM, MörgelinM, LinderA, et al. (2009) Treatment of invasive streptococcal infection with a peptide derived from human high-molecular-weight kininogen. Blood 114 : 444–451.
34. BrunoS, GrangeC, DeregibusMC, CalogeroRA, SaviozziS, et al. (2009) Mesenchymal stem cell-derived microvesicles protect against acute tubular injury. J Am Soc Nephrol 20 : 1053–1067.
35. BengtsonSH, PhagooSB, Norrby-TeglundA, PåhlmanL, MörgelinM, et al. (2006) Kinin receptor expression during Staphylococcus aureus infection. Blood 108 : 2055–2063.
36. BoberM, EnochssonC, CollinM, MörgelinM (2010) Collagen VI is a subepithelial adhesive target for human respiratory tract pathogens. J Innate Immun 2 : 160–166.
37. MalmströmJ, KarlssonC, NordenfeltP, OssolaR, WeisserH, et al. (2012) Streptococcus pyogenes in human plasma: adaptive mechanisms analyzed by mass spectrometry-based proteomics. J Biol Chem 287 : 1415–1425.
38. CraigR, BeavisRC (2003) A method for reducing the time required to match protein sequences with tandem mass spectra. Rapid Commun Mass Spectrom 17 : 2310–2316.
39. KellerA, EngJ, ZhangN, LiX-J, AebersoldR (2005) A uniform proteomics MS/MS analysis platform utilizing open XML file formats. Mol Syst Biol 1 : 2005.0017.
40. HodnikV, AnderluhG (2010) Capture of intact liposomes on biacore sensor chips for protein-membrane interaction studies. Methods Mol Biol 627 : 201–211.
Zvýšte si kvalifikáciu online z pohodlia domova
Nemáte účet? Registrujte sa
Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.
