Plasminogen activation by staphylokinase enhances local spreading of S. aureus in skin infections

BMC Microbiology

Peter Verhamme²  Roger H Lijnen²  Marc F Hoylaerts²  Bart De Geest²  Tao Jin³  Jakub Kwiecinksi³  Greetje Vande Velde¹  Jorien Claes²  Laurens Liesenborghs²  Thomas Vanassche²  Marijke Peetermans² 

[1] Biomedical MRI/Molecular Small Animal Imaging Center, KU Leuven, Herestraat 49, Leuven, Belgium;Center for Molecular and Vascular Biology, KU Leuven, Herestraat 49, Leuven, Belgium;Department of Rheumatology and Inflammation Research, University of Gothenburg, Guldhedsgatan 10, Gothenburg, Sweden

关键词: Skin infection;    Staphylokinase;    Staphylococcus aureus;    Plasminogen;    Matrix metalloproteinases;    Fibrinolysis;   

Others  :  1090660
DOI  :  10.1186/s12866-014-0310-7

 received in 2014-07-23, accepted in 2014-11-24,  发布年份 2014

【 摘 要 】

Background

Staphylococcus aureus (S. aureus) is a frequent cause of skin and soft tissue infections. A unique feature of S. aureus is the combined presence of coagulases that trigger fibrin formation and of the plasminogen activator staphylokinase (SAK). Whereas the importance of fibrin generation for S. aureus virulence has been established, the role of SAK remains unclear.

We studied the role of plasminogen activation by SAK in a skin infection model in mice and evaluated the impact of alpha-2-antiplasmin (α₂AP) deficiency on the spreading and proteolytic activity of S. aureus skin infections. The species-selectivity of SAK was overcome by adenoviral expression of human plasminogen. Bacterial spread and density was assessed non-invasively by imaging the bioluminescence of S. aureus Xen36.

Results

SAK-mediated plasmin activity increased the local invasiveness of S. aureus, leading to larger lesions with skin disruption as well as decreased bacterial clearance by the host. Even though fibrin and bacterial surfaces protected SAK-mediated plasmin activity from inhibition by α₂AP, the deficiency of α₂AP resulted in increased bacterial spreading. SAK-mediated plasmin also induced secondary activation of gelatinases, shown both in vitro and in lesions from the in vivo model.

Conclusion

SAK contributes to the phenotype of S. aureus skin infections by enhancing bacterial spreading as a result of fibrinolytic and proteolytic activation.

【 授权许可】

   
2014 Peetermans et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Ray GT, Suaya JA, Baxter R: Microbiology of skin and soft tissue infections in the age of community-acquired methicillin-resistant Staphylococcus aureus. Diagn Microbiol Infect Dis 2013, 76:24-30.
• [2]Sader HS, Farrell DJ, Jones RN: Antimicrobial susceptibility of Gram-positive cocci isolated from skin and skin-structure infections in European medical centres. Int J Antimicrob Agents 2010, 36:28-32.
• [3]Vanassche T, Verhaegen J, Peetermans WE, Van Ryn J, Cheng A, Schneewind O, Hoylaerts MF, Verhamme P: Inhibition of staphylothrombin by dabigatran reduces Staphylococcus aureus virulence. J Thromb Haemost 2011, 9:2436-2446.
• [4]Vanassche T, Peetermans M, Van Aelst L, Peetermans WE, Verhaegen J, Missiakas D, Schneewind O, Hoylaerts M, Verhamme P: The role of staphylothrombin-mediated fibrin deposition in catheter-related Staphylococcus aureus infections. J Infect Dis 2013, 208:92-100.
• [5]McAdow M, Kim HK, Dedent AC, Hendrickx AP, Schneewind O, Missiakas DM: Preventing Staphylococcus aureus sepsis through the inhibition of its agglutination in blood. PLoS Pathog 2011, 7:e1002307.
• [6]Sawai T, Tomono K, Yanagihara K, Yamamoto Y, Kaku M, Hirakata Y, Koga H, Tashiro T, Kohno S: 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.
• [7]Collen D: Staphylokinase: a potent, uniquely fibrin-selective thrombolytic agent. Nat Med 1998, 4:279-284.
• [8]Lijnen HR, Van Hoef B, De Cock F, Okada K, Ueshima S, Matsuo O, Collen D: On the mechanism of fibrin-specific plasminogen activation by staphylokinase. J Biol Chem 1991, 266:11826-11832.
• [9]Kwiecinski J, Josefsson E, Mitchell J, Higgins J, Magnusson M, Foster T, Jin T, Bokarewa M: Activation of plasminogen by staphylokinase reduces the severity of Staphylococcus aureus systemic infection. J Infect Dis 2010, 202:1041-1049.
• [10]Kwiecinski J, Jacobsson G, Karlsson M, Zhu X, Wang W, Bremell T, Josefsson E, Jin T: Staphylokinase Promotes the Establishment of Staphylococcus aureus Skin Infections While Decreasing Disease Severity. J Infect Dis 2013, 208:990-9.
• [11]Lijnen HR, Stassen JM, Vanlinthout I, Fukao H, Okada K, Matsuo O, Collen D: Comparative fibrinolytic properties of staphylokinase and streptokinase in animal models of venous thrombosis. Thromb Haemost 1991, 66:468-473.
• [12]Sun H, Ringdahl U, Homeister JW, Fay WP, Engleberg NC, Yang AY, Rozek LS, Wang X, Sjobring U, Ginsburg D: Plasminogen is a critical host pathogenicity factor for group A streptococcal infection. Science 2004, 305:1283-1286.
• [13]Khil J, Im M, Heath A, Ringdahl U, Mundada L, Cary Engleberg N, Fay WP: Plasminogen enhances virulence of group A streptococci by streptokinase-dependent and streptokinase-independent mechanisms. J Infect Dis 2003, 188:497-505.
• [14]Cook SM, Skora A, Walker MJ, Sanderson-Smith ML, McArthur JD: Site-restricted plasminogen activation mediated by group A streptococcal streptokinase variants. Biochem J 2014, 458:23-31.
• [15]Piechowicz L, Galin'ski J, Garbacz K, Haras K: Bacteriophage analysis of staphylokinase-negative Staphylococcus aureus strains isolated from people. J Basic Microbiol 2010, 50:557-561.
• [16]Jin T, Bokarewa M, McIntyre L, Tarkowski A, Corey GR, Reller LB, Fowler VG Jr: Fatal outcome of bacteraemic patients caused by infection with staphylokinase-deficient Staphylococcus aureus strains. J Med Microbiol 2003, 52:919-923.
• [17]Ruotsalainen E, Karden-Lilja M, Kuusela P, Vuopio-Varkila J, Virolainen-Julkunen A, Sarna S, Valtonen V, Jarvinen A: Methicillin-sensitive Staphylococcus aureus bacteraemia and endocarditis among injection drug users and nonaddicts: host factors, microbiological and serological characteristics. J Infect 2008, 56:249-256.
• [18]Luedicke C, Slickers P, Ehricht R, Monecke S: Molecular fingerprinting of Staphylococcus aureus from bone and joint infections. Eur J Clin Microbiol Infect Dis 2010, 29:457-463.
• [19]Monecke S, Luedicke C, Slickers P, Ehricht R: Molecular epidemiology of Staphylococcus aureus in asymptomatic carriers. Eur J Clin Microbiol Infect Dis 2009, 28:1159-1165.
• [20]Monecke S, Berger-Bachi B, Coombs G, Holmes A, Kay I, Kearns A, Linde HJ, O'Brien F, Slickers P, Ehricht R: Comparative genomics and DNA array-based genotyping of pandemic Staphylococcus aureus strains encoding Panton-Valentine leukocidin. Clin Microbiol Infect 2007, 13:236-249.
• [21]van Wamel WJ, Rooijakkers SH, Ruyken M, van Kessel KP, van Strijp JA: The innate immune modulators staphylococcal complement inhibitor and chemotaxis inhibitory protein of Staphylococcus aureus are located on beta-hemolysin-converting bacteriophages. J Bacteriol 2006, 188:1310-1315.
• [22]Humphreys H, Keane CT, Hone R, Pomeroy H, Russell RJ, Arbuthnott JP, Coleman DC: Enterotoxin production by Staphylococcus aureus isolates from cases of septicaemia and from healthy carriers. J Med Microbiol 1989, 28:163-172.
• [23]Katayama Y, Baba T, Sekine M, Fukuda M, Hiramatsu K: Beta-hemolysin promotes skin colonization by Staphylococcus aureus. J Bacteriol 2013, 195:1194-1203.
• [24]Resch G, Francois P, Morisset D, Stojanov M, Bonetti EJ, Schrenzel J, Sakwinska O, Moreillon P: Human-to-bovine jump of Staphylococcus aureus CC8 is associated with the loss of a beta-hemolysin converting prophage and the acquisition of a new staphylococcal cassette chromosome. PLoS ONE 2013, 8:e58187.
• [25]Gladysheva IP, Turner RB, Sazonova IY, Liu L, Reed GL: Coevolutionary patterns in plasminogen activation. Proc Natl Acad Sci U S A 2003, 100:9168-9172.
• [26]Kim SH, Chun HS, Han MH, Park NY, Suk K: A novel variant of staphylokinase gene from Staphylococcus aureus ATCC 29213. Thromb Res 1997, 87:387-395.
• [27]Sako T, Sawaki S, Sakurai T, Ito S, Yoshizawa Y, Kondo I: Cloning and expression of the staphylokinase gene of Staphylococcus aureus in Escherichia coli. Mol Gen Genet 1983, 190:271-277.
• [28]Behnke D, Gerlach D: Cloning and expression in Escherichia coli, Bacillus subtilis, and Streptococcus sanguis of a gene for staphylokinase–a bacterial plasminogen activator. Mol Gen Genet 1987, 210:528-534.
• [29]Coleman DC, Sullivan DJ, Russell RJ, Arbuthnott JP, Carey BF, Pomeroy HM: Staphylococcus aureus bacteriophages mediating the simultaneous lysogenic conversion of beta-lysin, staphylokinase and enterotoxin A: molecular mechanism of triple conversion. J Gen Microbiol 1989, 135:1679-1697.
• [30]Sumby P, Waldor MK: Transcription of the toxin genes present within the Staphylococcal phage phiSa3ms is intimately linked with the phage's life cycle. J Bacteriol 2003, 185:6841-6851.
• [31]Lijnen HR: Plasmin and matrix metalloproteinases in vascular remodeling. Thromb Haemost 2001, 86:324-333.
• [32]Collen D: Identification and some properties of a new fast-reacting plasmin inhibitor in human plasma. Eur J Biochem 1976, 69:209-216.
• [33]Molkanen T, Tyynela J, Helin J, Kalkkinen N, Kuusela P: Enhanced activation of bound plasminogen on Staphylococcus aureus by staphylokinase. FEBS Lett 2002, 517:72-78.
• [34]Furuya H, Ikeda R: Interaction of triosephosphate isomerase from Staphylococcus aureus with plasminogen. Microbiol Immunol 2011, 55:855-862.
• [35]Koch TK, Reuter M, Barthel D, Bohm S, van den Elsen J, Kraiczy P, Zipfel PF, Skerka C: Staphylococcus aureus proteins Sbi and Efb recruit human plasmin to degrade complement C3 and C3b. PLoS ONE 2012, 7:e47638.
• [36]Souza NM, Vieira ML, Alves IJ, de Morais ZM, Vasconcellos SA, Nascimento AL: Lsa30, a novel adhesin of Leptospira interrogans binds human plasminogen and the complement regulator C4bp. Microb Pathog 2012, 53:125-134.
• [37]Jin T, Bokarewa M, Foster T, Mitchell J, Higgins J, Tarkowski A: Staphylococcus aureus resists human defensins by production of staphylokinase, a novel bacterial evasion mechanism. J Immunol 2004, 172:1169-1176.
• [38]Rooijakkers SH, van Wamel WJ, Ruyken M, van Kessel KP, van Strijp JA: Anti-opsonic properties of staphylokinase. Microbes Infect 2005, 7:476-484.
• [39]Foster TJ, Hook M: Surface protein adhesins of Staphylococcus aureus. Trends Microbiol 2003, 48:1429-49.
• [40]Novick RP: Autoinduction and signal transduction in the regulation of staphylococcal virulence. Mol Microbiol 2003, 48:1429-1449.
• [41]Lijnen HR: Gene targeting in hemostasis. Alpha2-antiplasmin. Front Biosci 2001, 6:D239-247.
• [42]Feng Y, Jacobs F, Van Craeyveld E, Lievens J, Snoeys J, Van Linthout S, De Geest B: The impact of antigen expression in antigen-presenting cells on humoral immune responses against the transgene product. Gene Ther 2010, 17:288-293.
• [43]Van Linthout S, Lusky M, Collen D, De Geest B: Persistent hepatic expression of human apo A-I after transfer with a helper-virus independent adenoviral vector. Gene Ther 2002, 9:1520-1528.
• [44]Deutsch DG, Mertz ET: Plasminogen: purification from human plasma by affinity chromatography. Science 1970, 170:1095-1096.
• [45]Verhamme P, Goossens G, Maleux G, Collen D, Stas M: A dose-finding clinical trial of staphylokinase SY162 in patients with long-term venous access catheter thrombotic occlusion. J Thromb Thrombolysis 2007, 24:1-5.
• [46]Holvoet P, Lijnen HR, Collen D: A monoclonal antibody preventing binding of tissue-type plasminogen activator to fibrin: useful to monitor fibrinogen breakdown during t-PA infusion. Blood 1986, 67:1482-1487.
• [47]Van Hul M, Lijnen HR: Effect of weight loss on gelatinase levels in obese mice. Clin Exp Pharmacol Physiol 2011, 38:647-649.
• [48]Van Hul M, Lijnen HR: A functional role of gelatinase A in the development of nutritionally induced obesity in mice. J Thromb Haemost 2008, 6:1198-1206.