Fibrin formation by staphylothrombin facilitates Staphylococcus aureus-induced platelet aggregation

 

 Buy Article Permissions and Reprints

Summary

Interactions of Staphylococcus aureus (S. aureus) and platelets play an important role in the pathogenesis of intravascular infections such as infective endocarditis (IE). A typical feature of S. aureus is the ability to generate thrombin activity through the secretion of two prothrombin activating molecules, staphylocoagulase and von Willebrand factor-binding protein (vWbp), which bind to human prothrombin to form the enzymatically active staphylothrombin complex. The role of staphylothrombin in the interaction between S. aureus and platelets has not yet been studied. We found that in contrast with thrombin, staphylothrombin did not directly activate human platelets. However, the staphylothrombin-mediated conversion of fibrinogen to fibrin initiated platelet aggregation and secondary activation and facilitated S. aureus-platelet interactions. Both the genetic absence of staphylocoagulase and vWbp and pharmacological inhibition of staphylothrombin increased the lag time to aggregation, and reduced platelet trapping by S. aureus in high shear stress conditions. The combined inhibition of staphylothrombin and immunoglobulin binding to platelets completely abolished the ability of S. aureus to aggregate platelets in vitro. In conclusion, although staphylothrombin did not directly activate platelets, the formation of a fibrin scaffold facilitated bacteria-platelet interaction, and the inhibition of staphylothrombin resulted in a reduced activation of platelets by S. aureus.

• References

• 1 Cox D, Kerrigan SW, Watson SP. Platelets and the innate immune system: mechanisms of bacterial-induced platelet activation. J Thromb Haemost 2011; 9: 1097-1107.
  CrossrefPubMedGoogle Scholar
• 2 Verschoor A, Neuenhahn M, Navarini AA. et al. A platelet-mediated system for shuttling blood-borne bacteria to CD8alpha(+) dendritic cells depends on glycoprotein GPIb and complement C3. Nat Immunol 2011; 12: 1194-1201.
  CrossrefPubMedGoogle Scholar
• 3 Moreillon P, Que YA, Bayer AS. Pathogenesis of streptococcal and staphylococcal endocarditis. Infect Dis Clin North Am 2002; 16: 297-318.
  CrossrefPubMedGoogle Scholar
• 4 Kupferwasser LI, Yeaman MR, Nast CC. et al. Salicylic acid attenuates virulence in endovascular infections by targeting global regulatory pathways in Staphylococcus aureus. J Clin Invest 2003; 112: 222-233.
  CrossrefPubMedGoogle Scholar
• 5 Kupferwasser LI, Yeaman MR, Shapiro SM. et al. Acetylsalicylic acid reduces vegetation bacterial density, hematogenous bacterial dissemination, and frequency of embolic events in experimental Staphylococcus aureus endocarditis through antiplatelet and antibacterial effects. Circulation 1999; 99: 2791-2797.
  CrossrefPubMedGoogle Scholar
• 6 Kerrigan SW, Cox D. Platelet-bacterial interactions. Cell Mol Life Sci 2010; 67: 513-523.
  CrossrefPubMedGoogle Scholar
• 7 Kerrigan SW, Clarke N, Loughman A. et al. Molecular basis for Staphylococcus aureus-mediated platelet aggregate formation under arterial shear in vitro. Arterioscler Thromb Vasc Biol 2008; 28: 335-340.
  PubMedGoogle Scholar
• 8 Miajlovic H, Loughman A, Brennan M. et al. Both complement- and fibrinogen-dependent mechanisms contribute to platelet aggregation mediated by Staphylococcus aureus clumping factor B. Infect Immun 2007; 75: 3335-3343.
  CrossrefPubMedGoogle Scholar
• 9 Fitzgerald JR, Loughman A, Keane F. et al. Fibronectin-binding proteins of Staphylococcus aureus mediate activation of human platelets via fibrinogen and fibronectin bridges to integrin GPIIb/IIIa and IgG binding to the FcgammaRIIa receptor. Mol Microbiol 2006; 59: 212-230.
  CrossrefPubMedGoogle Scholar
• 10 Loughman A, Fitzgerald JR, Brennan MP. et al. Roles for fibrinogen, immunoglobulin and complement in platelet activation promoted by Staphylococcus aureus clumping factor A. Mol Microbiol 2005; 57: 804-818.
  CrossrefPubMedGoogle Scholar
• 11 Niemann S, Spehr N, Van Aken H. et al. Soluble fibrin is the main mediator of Staphylococcus aureus adhesion to platelets. Circulation 2004; 110: 193-200.
  CrossrefPubMedGoogle Scholar
• 12 Heilmann C, Niemann S, Sinha B. et al. Staphylococcus aureus fibronectin-binding protein (FnBP)-mediated adherence to platelets, and aggregation of platelets induced by FnBPA but not by FnBPB. J Infect Dis 2004; 190: 321-329.
  CrossrefPubMedGoogle Scholar
• 13 O'Brien L, Kerrigan SW, Kaw G. et al. Multiple mechanisms for the activation of human platelet aggregation by Staphylococcus aureus: roles for the clumping factors ClfA and ClfB, the serine-aspartate repeat protein SdrE and protein A. Mol Microbiol 2002; 44: 1033-1044.
  CrossrefPubMedGoogle Scholar
• 14 Bayer AS, Sullam PM, Ramos M. et al. Staphylococcus aureus induces platelet aggregation via a fibrinogen-dependent mechanism which is independent of principal platelet glycoprotein IIb/IIIa fibrinogen-binding domains. Infect Immun 1995; 63: 3634-3641.
  PubMedGoogle Scholar
• 15 Shannon O, Flock JI. Extracellular fibrinogen binding protein, Efb, from Staphylococcus aureus binds to platelets and inhibits platelet aggregation. Thromb Haemost 2004; 91: 779-789.
  Article in Thieme ConnectPubMedGoogle Scholar
• 16 Shannon O, Uekotter A, Flock JI. Extracellular fibrinogen binding protein, Efb, from Staphylococcus aureus as an antiplatelet agent in vivo. Thromb Haemost 2005; 93: 927-931.
  Article in Thieme ConnectPubMedGoogle Scholar
• 17 Loughman A, Fitzgerald JR, Brennan MP. et al. Roles for fibrinogen, immunoglobulin and complement in platelet activation promoted by Staphylococcus aureus clumping factor A. Mol Microbiol 2005; 57: 804-818.
  CrossrefPubMedGoogle Scholar
• 18 Friedrich R, Panizzi P, Fuentes-Prior P. et al. Staphylocoagulase is a prototype for the mechanism of cofactor-induced zymogen activation. Nature 2003; 425: 535-539.
  CrossrefPubMedGoogle Scholar
• 19 Kroh HK, Panizzi P, Bock PE. Von Willebrand factor-binding protein is a hysteretic conformational activator of prothrombin. Proc Natl Acad Sci USA 2009; 106: 7786-7791.
  CrossrefPubMedGoogle Scholar
• 20 Sawai T, Tomono K, Yanagihara K. et al. Role of coagulase in a murine model of hematogenous pulmonary infection induced by intravenous injection of Staphylococcus aureus enmeshed in agar beads. Infect Immun 1997; 65: 466-471.
  PubMedGoogle Scholar
• 21 McAdow M, Kim HK, Dedent AC. et al. Preventing Staphylococcus aureus Sepsis through the Inhibition of Its Agglutination in Blood. PLoS Pathog 2011; 7: e1002307.
  CrossrefPubMedGoogle Scholar
• 22 Cheng AG, McAdow M, Kim HK. et al. Contribution of coagulases towards Staphylococcus aureus disease and protective immunity. PLoS Pathog 2010; 6: e1001036.
  CrossrefPubMedGoogle Scholar
• 23 Vanassche T, Verhaegen J, Peetermans WE. et al. Inhibition of staphylothrombin by dabigatran reduces Staphylococcus aureus virulence. J Thromb Haemost 2011; 9: 2436-2446.
  CrossrefPubMedGoogle Scholar
• 24 Coughlin SR. Thrombin signalling and protease-activated receptors. Nature 2000; 407: 258-264.
  CrossrefPubMedGoogle Scholar
• 25 Vanassche T, Verhaegen J, Peetermans WE. et al. Dabigatran inhibits Staphylococcus aureus coagulase activity. J Clin Microbiol 2010; 48: 4248-4250.
  CrossrefPubMedGoogle Scholar
• 26 Panizzi P, Nahrendorf M, Figueiredo JL. et al. In vivo detection of Staphylococcus aureus endocarditis by targeting pathogen-specific prothrombin activation. Nat Med 2011; 17: 1142-1146.
  CrossrefPubMedGoogle Scholar
• 27 Yeaman MR, Puentes SM, Norman DC. et al. Partial characterization and staphylocidal activity of thrombin-induced platelet microbicidal protein. Infect Immun 1992; 60: 1202-1209.
  PubMedGoogle Scholar
• 28 Yeaman MR, Sullam PM, Dazin PF. et al. Platelet microbicidal protein alone and in combination with antibiotics reduces Staphylococcus aureus adherence to platelets in vitro. Infect Immun 1994; 62: 3416-3423.
  PubMedGoogle Scholar
• 29 Xiong YQ, Bayer AS, Yeaman MR. Inhibition of intracellular macromolecular synthesis in Staphylococcus aureus by thrombin-induced platelet microbicidal proteins. J Infect Dis 2002; 185: 348-356.
  CrossrefPubMedGoogle Scholar
• 30 Mercier RC, Dietz RM, Mazzola JL. et al. Beneficial influence of platelets on antibiotic efficacy in an in vitro model of Staphylococcus aureus-induced endocarditis. Antimicrob Agents Chemother 2004; 48: 2551-2557.
  CrossrefPubMedGoogle Scholar
• 31 Dhawan VK, Yeaman MR, Bayer AS. Influence of in vitro susceptibility phenotype against thrombin-induced platelet microbicidal protein on treatment and prophylaxis outcomes of experimental Staphylococcus aureus endocarditis. J Infect Dis 1999; 180: 1561-1568.
  CrossrefPubMedGoogle Scholar
• 32 Fowler Jr VG, McIntyre LM, Yeaman MR. et al. In vitro resistance to thrombin-induced platelet microbicidal protein in isolates of Staphylococcus aureus from endocarditis patients correlates with an intravascular device source. J Infect Dis 2000; 182: 1251-1254.
  CrossrefPubMedGoogle Scholar
• 33 Kupferwasser LI, Yeaman MR, Shapiro SM. et al. In vitro susceptibility to thrombin-induced platelet microbicidal protein is associated with reduced disease progression and complication rates in experimental Staphylococcus aureus endocarditis: microbiological, histopathologic, and echocardiographic analyses. Circulation 2002; 105: 746-752.
  CrossrefPubMedGoogle Scholar
• 34 Jarvis GE, Atkinson BT, Frampton J. et al. Thrombin-induced conversion of fibrinogen to fibrin results in rapid platelet trapping which is not dependent on platelet activation or GPIb. Br J Pharmacol 2003; 138: 574-583.
  CrossrefPubMedGoogle Scholar
• 35 Herrmann M, Lai QJ, Albrecht RM. et al. Adhesion of Staphylococcus aureus to surface-bound platelets: role of fibrinogen/fibrin and platelet integrins. J Infect Dis 1993; 167: 312-322.
  CrossrefPubMedGoogle Scholar
• 36 Jirouskova M, Dyr JE, Suttnar J. et al. Platelet adhesion to fibrinogen, fibrin monomer, and fibrin protofibrils in flowing blood -- the effect of fibrinogen immobilization and fibrin formation. Thromb Haemost 1997; 78: 1125-1131.
  Article in Thieme ConnectPubMedGoogle Scholar
• 37 Miajlovic H, Loughman A, Brennan M. et al. Both complement- and fibrinogen-dependent mechanisms contribute to platelet aggregation mediated by Staphylococcus aureus clumping factor B. Infect Immun 2007; 75: 3335-3343.
  CrossrefPubMedGoogle Scholar
• 38 George NP, Wei Q, Shin PK. et al. Staphylococcus aureus adhesion via Spa, ClfA, and SdrCDE to immobilized platelets demonstrates shear-dependent behavior. Arterioscler Thromb Vasc Biol 2006; 26: 2394-2400.
  CrossrefPubMedGoogle Scholar
• 39 Sjobring U, Ringdahl U, Ruggeri ZM. Induction of platelet thrombi by bacteria and antibodies. Blood 2002; 100: 4470-4477.
  CrossrefPubMedGoogle Scholar
• 40 Anderson CL, Chacko GW, Osborne JM. et al. The Fc receptor for immunoglobulin G (Fc gamma RII) on human platelets. Semin Thromb Hemost 1995; 21: 1-9.
  Article in Thieme ConnectPubMedGoogle Scholar
• 41 Dryla A, Prustomersky S, Gelbmann D. et al. Comparison of antibody repertoires against Staphylococcus aureus in healthy individuals and in acutely infected patients. Clin Diagn Lab Immunol 2005; 12: 387-398.
  PubMedGoogle Scholar
• 42 Vanassche T, Peetermans WE, Herregods MC. et al. Anti-thrombotic therapy in infective endocarditis. Expert Rev Cardiovasc Ther 2011; 9: 1203-1219.
  CrossrefPubMedGoogle Scholar

• Supplementary Material