Assessment of the Dual Role of Clumping Factor A in S. Aureus Adhesion to Endothelium in Absence and Presence of Plasma

Abstract

Adhesion of Staphylococcus aureus to endothelial cells (ECs) is paramount in infective endocarditis. Bacterial proteins such as clumping factor A (ClfA) and fibronectin binding protein A (FnbpA) mediate adhesion to EC surface molecules and (sub)endothelial matrix proteins including fibrinogen (Fg), fibrin, fibronectin (Fn) and von Willebrand factor (vWF). We studied the influence of shear flow and plasma on the binding of ClfA and FnbpA (including its sub-domains A, A¹⁶⁺, ABC, CD) to coverslip-coated vWF, Fg/fibrin, Fn or confluent ECs, making use of Lactococcus lactis, expressing these adhesins heterologously. Global adherence profiles were similar in static and flow conditions. In the absence of plasma, L. lactis-clfA binding to Fg increased with shear forces, whereas binding to fibrin did not. The degree of adhesion of L. lactis-fnbpA to EC-bound Fn and of L. lactis-clfA to EC-bound Fg, furthermore, was similar to that of L. lactis-clfA to coated vWF domain A1, in the presence of vWF-binding protein (vWbp). Yet, in plasma, L. lactis-clfA adherence to activated EC-vWF/vWbp dropped over 10 minutes by 80% due to vWF-hydrolysis by a disintegrin and metalloproteinase with thrombospondin type 1 motif, member 13 and that of L. lactis-fnbpA likewise by > 70% compared to the adhesion in absence of plasma. In contrast, plasma Fg supported high L. lactis-clfA binding to resting and activated ECs. Or, in plasma S. aureus adhesion to active endothelium occurs mainly via two complementary pathways: a rapid but short-lived vWF/vWbp pathway and a stable integrin-coupled Fg-pathway. Hence, the pharmacological inhibition of ClfA-Fg interactions may constitute a valuable additive treatment in infective endocarditis.

Keywords

infective endocarditis - clumping factor A - fibronectin binding proteins - S. aureus - von Willebrand factor - fibrinogen - fibronectin

• References

• 1 Lowy FD. Staphylococcus aureus infections. N Engl J Med 1998; 339 (08) 520-532
• 2 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 (01) 212-230
• 3 Heying R, van de Gevel J, Que YA, Piroth L, Moreillon P, Beekhuizen H. Contribution of (sub)domains of Staphylococcus aureus fibronectin-binding protein to the proinflammatory and procoagulant response of human vascular endothelial cells. Thromb Haemost 2009; 101 (03) 495-504
• 4 Massey RC, Kantzanou MN, Fowler T. , et al. Fibronectin-binding protein A of Staphylococcus aureus has multiple, substituting, binding regions that mediate adherence to fibronectin and invasion of endothelial cells. Cell Microbiol 2001; 3 (12) 839-851
• 5 Piroth L, Que YA, Widmer E. , et al. The fibrinogen- and fibronectin-binding domains of Staphylococcus aureus fibronectin-binding protein A synergistically promote endothelial invasion and experimental endocarditis. Infect Immun 2008; 76 (08) 3824-3831
• 6 Wann ER, Gurusiddappa S, Hook M. The fibronectin-binding MSCRAMM FnbpA of Staphylococcus aureus is a bifunctional protein that also binds to fibrinogen. J Biol Chem 2000; 275 (18) 13863-13871
• 7 Patti JM, Allen BL, McGavin MJ, Höök M. MSCRAMM-mediated adherence of microorganisms to host tissues. Annu Rev Microbiol 1994; 48: 585-617
• 8 McDevitt D, Nanavaty T, House-Pompeo K. , et al. Characterization of the interaction between the Staphylococcus aureus clumping factor (ClfA) and fibrinogen. Eur J Biochem 1997; 247 (01) 416-424
• 9 Siboo IR, Cheung AL, Bayer AS, Sullam PM. Clumping factor A mediates binding of Staphylococcus aureus to human platelets. Infect Immun 2001; 69 (05) 3120-3127
• 10 Allgayer S, Rosenbach G, Tavares CA, Polido WD. Periodontal ligament distraction: esthetics and occlusal stability at the 2-year follow-up. Am J Orthod Dentofacial Orthop 2013; 143 (04) 535-546
• 11 Foster TJ. The remarkably multifunctional fibronectin binding proteins of Staphylococcus aureus. Eur J Clin Microbiol Infect Dis 2016; 35 (12) 1923-1931
• 12 Mazmanian SK, Liu G, Jensen ER, Lenoy E, Schneewind O. Staphylococcus aureus sortase mutants defective in the display of surface proteins and in the pathogenesis of animal infections. Proc Natl Acad Sci U S A 2000; 97 (10) 5510-5515
• 13 Mazmanian SK, Liu G, Ton-That H, Schneewind O. Staphylococcus aureus sortase, an enzyme that anchors surface proteins to the cell wall. Science 1999; 285 (5428): 760-763
• 14 Schneewind O, Mihaylova-Petkov D, Model P. Cell wall sorting signals in surface proteins of gram-positive bacteria. EMBO J 1993; 12 (12) 4803-4811
• 15 Ton-That H, Liu G, Mazmanian SK, Faull KF, Schneewind O. Purification and characterization of sortase, the transpeptidase that cleaves surface proteins of Staphylococcus aureus at the LPXTG motif. Proc Natl Acad Sci U S A 1999; 96 (22) 12424-12429
• 16 Pappelbaum KI, Gorzelanny C, Grässle S. , et al. Ultralarge von Willebrand factor fibers mediate luminal Staphylococcus aureus adhesion to an intact endothelial cell layer under shear stress. Circulation 2013; 128 (01) 50-59
• 17 Claes J, Vanassche T, Peetermans M. , et al. Adhesion of Staphylococcus aureus to the vessel wall under flow is mediated by von Willebrand factor-binding protein. Blood 2014; 124 (10) 1669-1676
• 18 Schwarz-Linek U, Werner JM, Pickford AR. , et al. Pathogenic bacteria attach to human fibronectin through a tandem beta-zipper. Nature 2003; 423 (6936): 177-181
• 19 Que YA, Haefliger JA, Piroth L. , et al. Fibrinogen and fibronectin binding cooperate for valve infection and invasion in Staphylococcus aureus experimental endocarditis. J Exp Med 2005; 201 (10) 1627-1635
• 20 Sinha B, Francois P, Que YA. , et al. Heterologously expressed Staphylococcus aureus fibronectin-binding proteins are sufficient for invasion of host cells. Infect Immun 2000; 68 (12) 6871-6878
• 21 Strindhall J, Lindgren PE, Löfgren S, Kihlström E. Variations among clinical isolates of Staphylococcus aureus to induce expression of E-selectin and ICAM-1 in human endothelial cells. FEMS Immunol Med Microbiol 2002; 32 (03) 227-235
• 22 Peacock SJ, Foster TJ, Cameron BJ, Berendt AR. Bacterial fibronectin-binding proteins and endothelial cell surface fibronectin mediate adherence of Staphylococcus aureus to resting human endothelial cells. Microbiology 1999; 145 (Pt 12): 3477-3486
• 23 Barczyk M, Carracedo S, Gullberg D. Integrins. Cell Tissue Res 2010; 339 (01) 269-280
• 24 Que YA, François P, Haefliger JA, Entenza JM, Vaudaux P, Moreillon P. Reassessing the role of Staphylococcus aureus clumping factor and fibronectin-binding protein by expression in Lactococcus lactis. Infect Immun 2001; 69 (10) 6296-6302
• 25 Claes J, Liesenborghs L, Peetermans M. , et al. Clumping factor A, von Willebrand factor-binding protein and von Willebrand factor anchor Staphylococcus aureus to the vessel wall. J Thromb Haemost 2017; 15 (05) 1009-1019
• 26 Que YA, Haefliger JA, Francioli P, Moreillon P. Expression of Staphylococcus aureus clumping factor A in Lactococcus lactis subsp. cremoris using a new shuttle vector. Infect Immun 2000; 68 (06) 3516-3522
• 27 Claes J, Liesenborghs L, Lox M, Verhamme P, Vanassche T, Peetermans M. In vitro and in vivo model to study bacterial adhesion to the vessel wall under flow conditions. J Vis Exp 2015; (100) e52862
• 28 Theilmeier G, Lenaerts T, Remacle C, Collen D, Vermylen J, Hoylaerts MF. Circulating activated platelets assist THP-1 monocytoid/endothelial cell interaction under shear stress. Blood 1999; 94 (08) 2725-2734
• 29 Entenza JM, Moreillon P, Senn MM. , et al. Role of sigmaB in the expression of Staphylococcus aureus cell wall adhesins ClfA and FnbA and contribution to infectivity in a rat model of experimental endocarditis. Infect Immun 2005; 73 (02) 990-998
• 30 Vanassche T, Kauskot A, Verhaegen J. , et al. Fibrin formation by staphylothrombin facilitates Staphylococcus aureus-induced platelet aggregation. Thromb Haemost 2012; 107 (06) 1107-1121
• 31 McAdow M, Missiakas DM, Schneewind O. Staphylococcus aureus secretes coagulase and von Willebrand factor binding protein to modify the coagulation cascade and establish host infections. J Innate Immun 2012; 4 (02) 141-148
• 32 Veltrop MH, Beekhuizen H, Thompson J. Bacterial species- and strain-dependent induction of tissue factor in human vascular endothelial cells. Infect Immun 1999; 67 (11) 6130-6138
• 33 Kerrigan SW, Clarke N, Loughman A, Meade G, Foster TJ, Cox D. Molecular basis for Staphylococcus aureus-mediated platelet aggregate formation under arterial shear in vitro. Arterioscler Thromb Vasc Biol 2008; 28 (02) 335-340
• 34 Niemann S, Spehr N, Van Aken H. , et al. Soluble fibrin is the main mediator of Staphylococcus aureus adhesion to platelets. Circulation 2004; 110 (02) 193-200
• 35 Bjerketorp J, Jacobsson K, Frykberg L. The von Willebrand factor-binding protein (vWbp) of Staphylococcus aureus is a coagulase. FEMS Microbiol Lett 2004; 234 (02) 309-314
• 36 Thomer L, Schneewind O, Missiakas D. Multiple ligands of von Willebrand factor-binding protein (vWbp) promote Staphylococcus aureus clot formation in human plasma. J Biol Chem 2013; 288 (39) 28283-28292
• 37 Friedrich R, Panizzi P, Fuentes-Prior P. , et al. Staphylocoagulase is a prototype for the mechanism of cofactor-induced zymogen activation. Nature 2003; 425 (6957): 535-539
• 38 Kroh HK, Panizzi P, Bock PE. Von Willebrand factor-binding protein is a hysteretic conformational activator of prothrombin. Proc Natl Acad Sci U S A 2009; 106 (19) 7786-7791
• 39 Schneider H, Vogt M, Boekenkamp R. , et al. Melody transcatheter valve: Histopathology and clinical implications of nine explanted devices. Int J Cardiol 2015; 189: 124-131
• 40 Kremer Hovinga JA, Zeerleder S, Kessler P. , et al. ADAMTS-13, von Willebrand factor and related parameters in severe sepsis and septic shock. J Thromb Haemost 2007; 5 (11) 2284-2290
• 41 Aibar J, Castro P, Espinosa G. , et al. ADAMTS-13 in critically ill patients with septic syndromes and noninfectious systemic inflammatory response syndrome. Shock 2015; 43 (06) 556-562