Phagocytosis Escape by a Protein That Connects Complement and Coagulation Proteins at the Bacterial Surface

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Upon contact with human plasma, bacteria are rapidly recognized by the complement system that labels their surface for uptake and clearance by phagocytic cells. Staphylococcus aureus secretes the 16 kD Extracellular fibrinogen binding protein (Efb) that binds two different plasma proteins using separate domains: the Efb N-terminus binds to fibrinogen, while the C-terminus binds complement C3. In this study, we show that Efb blocks phagocytosis of S. aureus by human neutrophils. In vitro, we demonstrate that Efb blocks phagocytosis in plasma and in human whole blood. Using a mouse peritonitis model we show that Efb effectively blocks phagocytosis in vivo, either as a purified protein or when produced endogenously by S. aureus. Mutational analysis revealed that Efb requires both its fibrinogen and complement binding residues for phagocytic escape. Using confocal and transmission electron microscopy we show that Efb attracts fibrinogen to the surface of complement-labeled S. aureus generating a ‘capsule’-like shield. This thick layer of fibrinogen shields both surface-bound C3b and antibodies from recognition by phagocytic receptors. This information is critical for future vaccination attempts, since opsonizing antibodies may not function in the presence of Efb. Altogether we discover that Efb from S. aureus uniquely escapes phagocytosis by forming a bridge between a complement and coagulation protein.

Vyšlo v časopise: Phagocytosis Escape by a Protein That Connects Complement and Coagulation Proteins at the Bacterial Surface. PLoS Pathog 9(12): e32767. doi:10.1371/journal.ppat.1003816
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003816

Souhrn

Zdroje

1. NathanC (2006) Neutrophils and immunity: challenges and opportunities. Nat Rev Immunol 6 : 173–182.

2. GasqueP (2004) Complement: a unique innate immune sensor for danger signals. Mol Immunol 41 : 1089–1098.

3. RicklinD, HajishengallisG, YangK, LambrisJD (2010) Complement: a key system for immune surveillance and homeostasis. Nat Immunol 11 : 785–797.

4. GrosP, MilderFJ, JanssenBJC (2008) Complement driven by conformational changes. Nat Rev Immunol 8 : 48–58.

5. WalportMJ (2001) Complement. First of two parts. N Engl J Med 344 : 1058–1066.

6. LowyF (1998) Staphylococcus aureus infections. N Engl J Med 339 (8) 520–32.

7. LiM, DiepBA, VillaruzAE, BraughtonKR, JiangX, et al. (2009) Evolution of virulence in epidemic community-associated methicillin-resistant Staphylococcus aureus. Proceedings of the National Academy of Sciences 106 : 5883–5888.

8. SchafferAC, LeeJC (2008) Vaccination and passive immunisation against Staphylococcus aureus. Int J Antimicrob Agents 32 Suppl 1: S71–S78.

9. DeDentA, KimHK, MissiakasD, SchneewindO (2012) Exploring Staphylococcus aureus pathways to disease for vaccine development. Semin Immunopathol 34 : 317–333.

10. PalmaM, NozohoorS, SchenningsT, HeimdahlA, FlockJI (1996) Lack of the extracellular 19-kilodalton fibrinogen-binding protein from Staphylococcus aureus decreases virulence in experimental wound infection. Infection and Immunity 64 : 5284–5289.

11. SchenningsT, FarneboF, SzekelyL, FlockJ-I (2012) Protective immunization against Staphylococcus aureus infection in a novel experimental wound model in mice. APMIS 120 : 786–793.

12. JongeriusI, KöhlJ, PandeyMK, RuykenM, van KesselKPM, et al. (2007) Staphylococcal complement evasion by various convertase-blocking molecules. J Exp Med 204 : 2461–2471.

13. KoY-P, LiangX, SmithCW, DegenJL, HöökM (2011) Binding of Efb from Staphylococcus aureus to fibrinogen blocks neutrophil adherence. Journal of Biological Chemistry 286 : 9865–9874.

14. HammelM, SfyroeraG, RicklinD, MagottiP, LambrisJD, et al. (2007) A structural basis for complement inhibition by Staphylococcus aureus. Nat Immunol 8 : 430–437.

15. LeeLYL, HöökM, HavilandD, WetselRA, YonterEO, et al. (2004) Inhibition of complement activation by a secreted Staphylococcus aureus protein. J Infect Dis 190 : 571–579.

16. PalmaM, ShannonO, QuezadaHC, BergA, FlockJ-I (2001) Extracellular fibrinogen-binding protein, Efb, from Staphylococcus aureus blocks platelet aggregation due to its binding to the alpha-chain. J Biol Chem 276 : 31691–31697.

17. ShannonO, FlockJ-I (2004) Extracellular fibrinogen binding protein, Efb, from Staphylococcus aureus binds to platelets and inhibits platelet aggregation. Thromb Haemost 91 (4) 779–89.

18. KochTK, ReuterM, BarthelD, BöhmS, van den ElsenJ, et al. (2012) Staphylococcus aureus proteins Sbi and Efb recruit human plasmin to degrade complement C3 and C3b. PLoS ONE 7 (10) e47638.

19. PalmaM, WadeD, FlockM, FlockJ-I (1998) Multiple binding sites in the interaction between an extracellular fibrinogen-binding protein from Staphylococcus aureus and fibrinogen. J Biol Chem 273 : 13177–13181.

20. PlowEF, HaasTA, ZhangL, LoftusJ, SmithJW (2000) Ligand binding to integrins. J Biol Chem 275 : 21785–21788.

21. FlickMJ, DuX, WitteDP, JirouskováM, SolovievDA, et al. (2004) Leukocyte engagement of fibrin(ogen) via the integrin receptor alphaMbeta2/Mac-1 is critical for host inflammatory response in vivo. J Clin Invest 113 : 1596–1606.

22. ThakkerM, ParkJS, CareyV, LeeJC (1998) Staphylococcus aureus serotype 5 capsular polysaccharide is antiphagocytic and enhances bacterial virulence in a murine bacteremia model. Infection and Immunity 66 : 5183–5189.

23. JongeriusI, Köckritz-Blickwede vonM, HorsburghMJ, RuykenM, NizetV, et al. (2012) Staphylococcus aureus Virulence Is Enhanced by Secreted Factors That Block Innate Immune Defenses. Journal of Innate Immunity 4 (3) 301–311.

24. PantrangiM, SinghVK, WolzC, ShuklaSK (2010) Staphylococcal superantigen-like genes, ssl5 and ssl8, are positively regulated by Sae and negatively by Agr in the Newman strain. FEMS Microbiology Letters 308 : 175–184.

25. VoyichJM, VuongC, (null), NygaardTK, (null), et al. (2009) The SaeR/S gene regulatory system is essential for innate immune evasion by Staphylococcus aureus. J Infect Dis 199 : 1698–1706.

26. LoofTG, MorgelinM, JohanssonL, OehmckeS, OlinAI, et al. (2011) Coagulation, an ancestral serine protease cascade, exerts a novel function in early immune defense. Blood 118 : 2589–2598.

27. FrickI-M, AkessonP, HerwaldH, MörgelinM, MalmstenM, et al. (2006) The contact system–a novel branch of innate immunity generating antibacterial peptides. EMBO J 25 : 5569–5578.

28. MattssonE, HerwaldH, CramerH, PerssonK, SjöbringU, et al. (2001) Staphylococcus aureus induces release of bradykinin in human plasma. Infection and Immunity 69 : 3877–3882.

29. NordenfeltP, WaldemarsonS, LinderA, MorgelinM, KarlssonC, et al. (2012) Antibody orientation at bacterial surfaces is related to invasive infection. Journal of Experimental Medicine 209 (13) 2367–81.

30. McCarthyAJ, LindsayJA (2010) Genetic variation in Staphylococcus aureus surface and immune evasion genes is lineage associated: implications for vaccine design and host-pathogen interactions. BMC Microbiol 10 : 173.

31. McAdowM, MissiakasDM, SchneewindO (2012) Staphylococcus aureus secretes coagulase and von Willebrand factor binding protein to modify the coagulation cascade and establish host infections. Journal of Innate Immunity 4 : 141–148.

32. ChengAG, McAdowM, KimHK, BaeT, MissiakasDM, et al. (2010) Contribution of coagulases towards Staphylococcus aureus disease and protective immunity. PLoS Pathog 6: e1001036.

33. NygaardTK, PallisterKB, RuzevichP, GriffithS, VuongC, et al. (2010) SaeR binds a consensus sequence within virulence gene promoters to advance USA300 pathogenesis. J Infect Dis 201 : 241–254.

34. ChavakisT, WiechmannK, PreissnerKT, HerrmannM (2005) Staphylococcus aureus interactions with the endothelium: the role of bacterial “secretable expanded repertoire adhesive molecules” (SERAM) in disturbing host defense systems. Thromb Haemost 94 : 278–285.

35. ChengAG, KimHK, BurtsML, KrauszT, SchneewindO, et al. (2009) Genetic requirements for Staphylococcus aureus abscess formation and persistence in host tissues. The FASEB Journal 23 : 3393–3404.

36. GuggenbergerC, WolzC, MorrisseyJA, HeesemannJ (2012) Two distinct coagulase-dependent barriers protect Staphylococcus aureus from neutrophils in a three dimensional in vitro infection model. PLoS Pathog 8: e1002434.

37. PangYY, SchwartzJ, ThoendelM, AckermannLW, HorswillAR, et al. (2010) agr-Dependent interactions of Staphylococcus aureus USA300 with human polymorphonuclear neutrophils. Journal of Innate Immunity 2 : 546–559.

38. RooijakkersSHM, WuJ, RuykenM, van DomselaarR, PlankenKL, et al. (2009) Structural and functional implications of the alternative complement pathway C3 convertase stabilized by a staphylococcal inhibitor. Nat Immunol 10 : 721–727.

39. EverseSJ, PelletierH, DoolittleRF (1995) Crystallization of fragment D from human fibrinogen. Protein Sci 4 : 1013–1016.

40. MollnesTE, BrekkeO-L, FungM, FureH, ChristiansenD, et al. (2002) Essential role of the C5a receptor in E coli-induced oxidative burst and phagocytosis revealed by a novel lepirudin-based human whole blood model of inflammation. Blood 100 : 1869–1877.

41. BestebroerJ, PoppelierMJJG, UlfmanLH, LentingPJ, DenisCV, et al. (2007) Staphylococcal superantigen-like 5 binds PSGL-1 and inhibits P-selectin-mediated neutrophil rolling. Blood 109 : 2936–2943.

42. HorsburghMJ, AishJL, WhiteIJ, ShawL, LithgowJK, et al. (2002) sigmaB modulates virulence determinant expression and stress resistance: characterization of a functional rsbU strain derived from Staphylococcus aureus 8325-4. J Bacteriol 184 : 5457–5467.

43. SlotJW, GeuzeHJ (2007) Cryosectioning and immunolabeling. Nat Protoc 2 : 2480–2491.