Preventing Sepsis through the Inhibition of Its Agglutination in Blood


Staphylococcus aureus infection is a frequent cause of sepsis in humans, a disease associated with high mortality and without specific intervention. When suspended in human or animal plasma, staphylococci are known to agglutinate, however the bacterial factors responsible for agglutination and their possible contribution to disease pathogenesis have not yet been revealed. Using a mouse model for S. aureus sepsis, we report here that staphylococcal agglutination in blood was associated with a lethal outcome of this disease. Three secreted products of staphylococci - coagulase (Coa), von Willebrand factor binding protein (vWbp) and clumping factor (ClfA) – were required for agglutination. Coa and vWbp activate prothrombin to cleave fibrinogen, whereas ClfA allowed staphylococci to associate with the resulting fibrin cables. All three virulence genes promoted the formation of thromboembolic lesions in heart tissues. S. aureus agglutination could be disrupted and the lethal outcome of sepsis could be prevented by combining dabigatran-etexilate treatment, which blocked Coa and vWbp activity, with antibodies specific for ClfA. Together these results suggest that the combined administration of direct thrombin inhibitors and ClfA-antibodies that block S. aureus agglutination with fibrin may be useful for the prevention of staphylococcal sepsis in humans.


Vyšlo v časopise: Preventing Sepsis through the Inhibition of Its Agglutination in Blood. PLoS Pathog 7(10): e32767. doi:10.1371/journal.ppat.1002307
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002307

Souhrn

Staphylococcus aureus infection is a frequent cause of sepsis in humans, a disease associated with high mortality and without specific intervention. When suspended in human or animal plasma, staphylococci are known to agglutinate, however the bacterial factors responsible for agglutination and their possible contribution to disease pathogenesis have not yet been revealed. Using a mouse model for S. aureus sepsis, we report here that staphylococcal agglutination in blood was associated with a lethal outcome of this disease. Three secreted products of staphylococci - coagulase (Coa), von Willebrand factor binding protein (vWbp) and clumping factor (ClfA) – were required for agglutination. Coa and vWbp activate prothrombin to cleave fibrinogen, whereas ClfA allowed staphylococci to associate with the resulting fibrin cables. All three virulence genes promoted the formation of thromboembolic lesions in heart tissues. S. aureus agglutination could be disrupted and the lethal outcome of sepsis could be prevented by combining dabigatran-etexilate treatment, which blocked Coa and vWbp activity, with antibodies specific for ClfA. Together these results suggest that the combined administration of direct thrombin inhibitors and ClfA-antibodies that block S. aureus agglutination with fibrin may be useful for the prevention of staphylococcal sepsis in humans.


Zdroje

1. LowyFD 1998 Staphylococcus aureus infections. New Engl J Med 339 520 532

2. KlevensRMMorrisonMANadleJPetitSGershmanK 2007 Invasive methicillin-resistant Staphylococcus aureus infections in the United States. JAMA 298 1763 1771

3. DeLeoFRChambersHF 2009 Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol 7 629 641

4. FowlerVGJMiroJMHoenBCabellCHAbrutynE 2005 Staphylococcus aureus endocarditis: a consequence of medical progress. JAMA 293 3012 3021

5. DeLeoFROttoMKreiswirthBNChambersHF 2010 Community-associated meticillin-resistant Staphylococcus aureus. Lancet 375 1557 1568

6. FosterTJ 2005 Immune evasion by staphylococci. Nat Rev Microbiol 3 948 958

7. WalshEJMiajlovicHGorkunOVFosterTJ 2008 Identification of the Staphylococcus aureus MSCRAMM clumping factor B (ClfB) binding site in the alphaC-domain of human fibrinogen. Microbiology 154 550 558

8. ChengAGDeDentACSchneewindOMissiakasDM 2011 A play in four acts: Staphylococcus aureus abscess formation. Trends Microbiol 19 225 232

9. DoolittleRF 2003 Structural basis of the fibrinogen-fibrin transformation: contributions from X-ray crystallography. Blood Rev 17 33 41

10. PanizziPFriedrichRFuentes-PriorPRichterKBockPE 2006 Fibrinogen substrate recognition by staphylocoagulase (pro)thrombin complexes. J Biol Chem 281 1179 1187

11. FriedrichRPanizziPFuentes-PriorPRichterKVerhammeI 2003 Staphylocoagulase is a prototype for the mechanism of cofactor-induced zymogen activation. Nature 425 535 539

12. ChengAGMcAdowMKimHKBaeTMissiakasDM 2010 Contribution of coagulases towards Staphylococcus aureus disease and protective immunity. PLoS Pathog 6 e1001036

13. LoebL 1903 The influence of certain bacteria on the coagulation of blood. J Med Res 10 407

14. KolleWOttoR 1902 Die Differenzierung der Staphylokokken mittelst der Agglutination. Z Hygiene 41 369 379

15. McDevittDFrancoisPVaudauxPFosterTJ 1994 Molecular characterization of the clumping factor (fibrinogen receptor) of Staphylococcus aureus. Mol Microbiol 11 237 248

16. HawigerJTimmonsSStrongDDCottrellBARileyM 1982 Identification of a region of human fibrinogen interacting with staphylococcal clumping factor. Biochemistry 21 1407 1413

17. McDevittDFrancoisPVaudauxPFosterTJ 1995 Identification of the ligand-binding domain of the surface-located fibrinogen receptor (clumping factor) of Staphylococcus aureus. Mol Microbiol 16 895 907

18. McDevittDNanavatyTHouse-PompeoKBellETurnerN 1997 Characterization of the interaction between the Staphylococcus aureus clumping factor (ClfA) and fibrinogen. Eur J Biochem 247 416 424

19. StrongDDLaudanoAPHawigerJDoolittleRF 1982 Isolation, characterization and synthesis of peptides from human fibrinogen that block the staphylococcal clumping reaction and construction of a synthetic clumping particle. Biochemistry 21 1414 1420

20. GaneshVKRiveraJJSmedsEKoY-PBowdenMG 2008 A structural model of the Staphylococcus aureus ClfA-fibrinogen interaction opens new avenues for the design of anti-staphylococcal therapeutics. PLoS Pathog 4 e1000226

21. JosefssonEHartfordOO'BrienLPattiJMFosterTJ 2001 Protection against experimental Staphylococcus aureus arthritis by vaccination with clumping factor A, a novel virulence determinant. J Infect Dis 184 1572 1580

22. MoreillonPEntenzaJMFrancioliPMcDevittDFosterTJ 1995 Role of Staphylococcus aureus coagulase and clumping factor in pathogenesis of experimental endocarditis. Infect Immun 63 4738 4743

23. HairPSEchagueCGShollAMWatkinsJAGeogheganJA 2010 Clumping factor A interaction with complement factor I increases C3b cleavage on the bacterial surface of Staphylococcus aureus and decreases complement-mediated phagocytosis. Infect Immun 78 1717 1727

24. HallAEDomanskiPJPatelPRVernachioJHSyribeysPJ 2003 Characterization of a protective monoclonal antibody recognizing Staphylococcus aureus MSCRAMM protein clumping factor A. Infect Immun 71 6864 6870

25. WeemsJJJrSteinbergJPFillerSBaddleyJWCoreyGR 2006 Phase II, randomized, double-blind, multicenter study comparing the safety and pharmacokinetics of Tefibazumab to placebo for treatment of Staphylococcus aureus bacteremia. Antimicrob Agents Chemother 50 2751 2755

26. Birch-HirschfeldL 1934 Über die Agglutination von Staphylokokken durch Bestandteile des Säugetierblutplasmas. Klinische Woschenschrift 13 331

27. KimHKDeDentAChengAGMcAdowMBagnoliF 2010 IsdA and IsdB antibodies protect mice against Staphylococcus aureus abscess formation and lethal challenge. Vaccine 28 6382 6392

28. MazmanianSKLiuGTon-ThatHSchneewindO 1999 Staphylococcus aureus sortase, an enzyme that anchors surface proteins to the cell wall. Science 285 760 763

29. BaeTBangerAKWallaceAGlassEMAslundF 2004 Staphylococcus aureus virulence genes identified by bursa aurealis mutagenesis and nematode killing. Proc Natl Acad Sci U S A 101 12312 12317

30. KaidaSMiyataTYoshizawaYKawabataSMoritaT 1987 Nucleotide sequence of the staphylocoagulase gene: its unique COOH-terminal 8 tandem repeats. J Biochem 102 1177 1186

31. BjerketorpJNilssonMLjunghAFlockJIJacobssonK 2002 A novel von Willebrand factor binding protein expressed by Staphylococcus aureus. Microbiology 148 2037 2044

32. PalmaMNozohoorSSchenningsTHeimdahlAFlockJI 1996 Lack of the extracellular 19-kilodalton fibrinogen-binding protein from Staphylococcus aureus decreases virulence in experimental wound infection. Infect Immun 64 5284 5289

33. PalmaMWadeDFlockMFlockJI 1998 Multiple binding sites in the interaction between an extracellular fibrinogen-binding protein from Staphylococcus aureus and fibrinogen. J Biol Chem 273 13177 13181

34. Ní EidhinDPerkinsSFrancoisPVaudauxPHöökM 1998 Clumping factor B (ClfB), a new surface-located fibrinogen-binding adhesin of Staphylococcus aureus. Mol Microbiol 30 245 257

35. HawigerJHammondDKTimmonsS 1975 Human fibrinogen possesses binding sites for staphylococci on Aalpha and Bbeta polypeptide chains. Nature 258 643 645

36. Stranger-JonesYKBaeTSchneewindO 2006 Vaccine assembly from surface proteins of Staphylococcus aureus. Proc Nat Acad Sci U S A 103 16942 16947

37. Hijikata-OkunomiyaAKataokaN 2003 Argatroban inhibits staphylothrombin. J Thromb Haemost 1 2060 2061

38. VanasscheTVerhaegenJPeetermansWEHoylaertsMFVerhammeP 2010 Dabigatran inhibits Staphylococcus aureus coagulase activity. J Clin Microbiol 48 4248 4250

39. HaulNHNarHPriepkeHRiesUStassenJM 2002 Structure-based design of novel potent nonpeptide thrombin inhibitors. J Med Chem 45 1757 1766

40. BabaTTakeuchiFKurodaMYuzawaHAokiK 2002 Genome and virulence determinants of high-virulence community acquired MRSA. Lancet 359 1819 1827

41. KurodaMOhtaTUchiyamaIBabaTYuzawaH 2001 Whole genome sequencing of meticillin-resistant Staphylococcus aureus. Lancet 357 1225 1240

42. Stearns-KurosawaDJOsuchowskiMFValentineCKurosawaSRemickDG 2010 The pathogenesis of sepsis. Annu Rev Pathol Mech Dis 6 19 48

43. WarrenHSSuffrediniAFEichackerPQMunfordRS 2002 Risks and benefits of activated protein C treatment for severe sepsis. N Engl J Med 347 1027 1030

44. JaimesFDe La RosaGMoralesCFortichFArangoC 2009 Unfractioned heparin for treatment of sepsis: A randomized clinical trial (The HETRASE Study). Crit Care Med 37 1185 1196

45. Di NisioMMiddeldorpSBullerHR 2005 Direct thrombin inhibitors. N Engl J Med 353 1028 1040

46. MarlarRAKleissAJGriffinJH 1981 Human protein C: inactivation of factors V and VIII in plasma by the activated molecule. Ann NY Acad Sci 370 303 310

47. BernardGRVincentJLLaterrePFLaRosaSPDhainautJF 2001 Efficacy and safety of recombinant human activated protein C for severe sepsis. N Engl J Med 344 699 709

48. LevyMMDellingerRPTownsendSRLinde-ZwirbleWTMarshallJC 2010 The Surviving Sepsis Campaign: results of an international guideline-based performance improvement program targeting severe sepsis. Intensive Care Med 36 222 231

49. TaylorFBJChangAEsmonCTD'AngeloAVigano-D'AngeloS 1987 Protein C prevents the coagulopathic and lethal effects of Escherichia coli infusion in the baboon. J Clin Invest 79 918 925

50. MartinGSManninoDMEatonSMossM 2003 The epidemiology of sepsis in the United States from 1979 through 2000. N Engl J Med 348 1546 1554

51. LiuCBayerASCosgroveSEDaumRSFridkinSK 2011 Clinical practice guidelines by the infectious diseases society of america for the treatment of methicillin-resistant Staphylococcus aureus infections in adults and children: executive summary. Clin Infect Dis 52 285 292

52. WalshCT 1993 Vancomycin resistance: decoding the molecular logic. Science 261 308 309

53. FowlerVGJrBoucherHWCoreyGRAbrutynEKarchmerAW 2006 Daptomycin versus standard therapy for bacteremia and edocarditis caused by Staphylococcus aureus. N Engl J Med 355 653 665

54. KlevensRMEdwardsJRGaynesRPSystemNNIS 2008 The impact of antimicrobial-resistant, health care-associated infections on mortality in the United States. Clin Infect Dis 47 927 930

55. McCarthyAJLindsayJA 2010 Genetic variation in Staphylococcus aureus surface and immune evasion genes is lineage associated: implications for vaccine design and host-pathogen interactions. BMC Microbiology 10 173

56. ColemanDCSullivanDJRussellRJArbuthnottJPCareyBF 1989 Staphylococcus aureus bacteriophages mediating the simultaneous lysogenic conversion of beta-lysin, staphylokinase and enterotoxin A: a molecular mechanism of triple conversion. J Gen Microbiol 135 1679 1697

57. LijnenHRVan HoefBDe CockFOkadaKUeshimaS 1991 On the mechanism of fibrin-specific plasminogen activation by staphylokinase. J Biol Chem 266 11826 11832

58. MölkänenTTyyneläJHelinJKalkkinenNKuuselaP 2002 Enhanced activation of bound plasminogen on Staphylococcus aureus by staphylokinase. FEBS Lett 517 72 78

59. JinTBokarewaMFosterTMitchellJHigginsJ 2004 Staphylococcus aureus resists human defensins by production of staphylokinase, a novel bacterial evasion mechanism. J Immunol 172 1169 1176

60. MazmanianSKLiuGJensenERLenoyESchneewindO 2000 Staphylococcus aureus mutants defective in the display of surface proteins and in the pathogenesis of animal infections. Proc Natl Acad Sci U S A 97 5510 5515

61. BabaTBaeTSchneewindOTakeuchiFHiramatsuK 2007 Genome sequence of Staphylococcus aureus strain Newman and comparative analysis of staphylococcal genomes. J Bacteriol 190 300 310

62. DiepBAGillSRChangRFPhanTHChenJH 2006 Complete genome sequence of USA300, an epidemic clone of community-acquired meticillin-resistant Staphylococcus aureus. Lancet 367 731 739

63. DonahueJPPatelHAndersonWFHawigerJ 1994 Three-dimensional structure of the platelet integrin recognition segment of the fibrinogen gamma chain obtained by carrier protein-driven characterization. Proc Natl Acad Sci U S A 91 12178 12182

64. WareSDonahueJPHawigerJAndersonWF 1999 Structure of the fibrinogen gamma-chain integrin binding and factor XIIIa cross-linking sites obtained through carrier protein driven crystallization. Protein Sci 8 2663 2671

65. WolbergAS 2007 Thrombin generation and fibrin structure. Blood Rev 21 131 142

66. SchneewindOMihaylova-PetkovDModelP 1993 Cell wall sorting signals in surface protein of Gram-positive bacteria. EMBO 12 4803 4811

67. DeDentACMissiakasDMSchneewindO 2008 Signal peptides direct surface proteins to two distinct envelope locations of Staphylococcus aureus. EMBO J 27 2656 2668

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2011 Číslo 10
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Eozinofilní granulomatóza s polyangiitidou
nový kurz
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa