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Thromb Haemost 2006; 95(05): 836-849
DOI: 10.1160/TH05-08-0534
Wound Healing and Inflammation/Infection
Schattauer GmbH

Group B streptococcus isolates from septic patients and healthy carriers differentially activate platelet signaling cascades

Celine Siauw   *
1   University Children’s Hospital, University of Wuerzburg, Wuerzburg, Germany
,
Anna Kobsar   *
3   Institute of Clinical Biochemistry and Pathobiochemistry/ Central Laboratory, University of Wuerzburg, Wuerzburg, Germany
,
Catharina Dornieden
3   Institute of Clinical Biochemistry and Pathobiochemistry/ Central Laboratory, University of Wuerzburg, Wuerzburg, Germany
,
Claudia Beyrich
3   Institute of Clinical Biochemistry and Pathobiochemistry/ Central Laboratory, University of Wuerzburg, Wuerzburg, Germany
,
Birgitta Schinke
3   Institute of Clinical Biochemistry and Pathobiochemistry/ Central Laboratory, University of Wuerzburg, Wuerzburg, Germany
,
Alexandra Schubert-Unkmeir
2   Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany
,
Marianne Abele-Horn
2   Institute of Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany
,
Christian P. Speer
1   University Children’s Hospital, University of Wuerzburg, Wuerzburg, Germany
,
Martin Eigenthaler
3   Institute of Clinical Biochemistry and Pathobiochemistry/ Central Laboratory, University of Wuerzburg, Wuerzburg, Germany
› Author AffiliationsFinancial support: The authors were supported by a grant from the “Interdisziplinäres Zentrum für Klinische Forschung” (IZKF grant A33) of the University of Wuerzburg.
Further Information

Publication History

Received 01 August 2005

Accepted after resubmission 25 March 2006

Publication Date:
01 December 2017 (online)

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Summary

Infection with group B streptococcus (GBS) is the most common cause of early onset neonatal sepsis in many countries, leading to neonatal morbidity and mortality.There is much evidence fora direct involvement of platelets in the pathogenesis of inflammation and sepsis. Several bacteria are known to directly interact with platelets leading to activation and aggregation,a phenomenon also observed with GBS. Here, we demonstrate that GBS rapidly bound to platelets; however, only strains isolated from septic patients bound fibrinogen on their surface and induced platelet thromboxane synthesis, platelet aggregation, and P-selectin (CD62P) expression. In contrast, GBS strains isolated from healthy newborns or healthy pregnant women induced only shape change, but not platelet thromboxane synthesis, platelet aggregation, or CD62P expression. All GBS strains investigated were able to activate FcγRIIA receptor signaling pathways including phospholipase C gamma2 (PLCγ2), as well as calcium/calmodulin-dependent myosin kinase II (CaMKII) and phosphorylation of myosin light chain (MLC). In contrast, protein kinase C (PKC) was exclusively activated by GBS strains isolated from septic patients, and p38 mitogen activated protein kinase (p38 MAP kinase) was preferentially activated by septic GBS strains. Furthermore, stress signaling kinase SEK1/MKK4 and focal adhesion kinase (FAK) were activated by all tested GBS strains in a FcγRIIA-independent way.This study demonstrates that septic, but not colonizing, GBS strains bind fibrinogen on their surface, and that septic GBS strains influence platelet function not only via the FcγRIIA receptor, but also via pathways distinct from IgG-mediated signalling. These mechanisms lead to platelet aggregation and secretion, thereby possibly modulating the pathophysiologic course of GBS infections.

Keywords

GroupB streptococcus - platelets - FcγRIIA - MAPK - PKC

* Both authors contributed equally.


 

  • References

  • 1 Luck S, Torny M, d’Agapeyeff K. et al. Estimated early-onset group B streptococcal neonatal disease. Lancet 2003; 361: 1953-4.
    CrossrefPubMedGoogle Scholar
  • 2 Vincent JL, Yagushi A, Pradier O. Platelet function in sepsis. Crit Care Med 2002; 30: S313-7.
    CrossrefPubMedGoogle Scholar
  • 3 Usui Y, Ohshima Y, Yoshida K. Platelet aggregation by group B streptococci. J Gen Microbiol 1987; 133: 1593-600.
    PubMedGoogle Scholar
  • 4 Lorant DE, Topham MK, Whatley RE. et al. Inflammatory roles of P-selectin. J Clin Invest 1993; 92: 559-70.
    CrossrefPubMedGoogle Scholar
  • 5 Anderson GP, Anderson CL. Signal transduction by the platelet Fcγ receptor. Blood 1990; 76: 1165-72.
    PubMedGoogle Scholar
  • 6 Gratacap MP, Payrastre B, Viala C. et al. Phosphatidylinositol 3,4,5-trisphosphate-dependent stimulation of phospholipase C-gamma2 is an early key event in FcgammaRIIA-mediated activation of human platelets. J Biol Chem 1998; 273: 24314-21.
    CrossrefPubMedGoogle Scholar
  • 7 Daniel JL, Molish IR, Rigmaiden M. et al. Evidence fora role of myosin phosphorylation in the initiation of the platelet shape change response. J Biol Chem 1984; 259: 9826-31.
    PubMedGoogle Scholar
  • 8 Rink TJ, Smith SW, Tsien RY. Cytoplasmic free Ca2+ in human platelets: Ca2+ thresholds and Ca-independent activation for shape-change and secretion. FEBS Lett 1982; 148: 21-6.
    CrossrefPubMedGoogle Scholar
  • 9 Rhee SG, Bae YS. Regulation of phosphoinositidespecific phospholipase C isozymes. J Biol Chem 1997; 272: 15045-8.
    CrossrefPubMedGoogle Scholar
  • 10 Dandona P, Thusu K, Khurana U. et al. Calcium, calmodulin and protein kinase C dependence of platelet shape change. Thromb Res 1996; 81: 163-75.
    CrossrefPubMedGoogle Scholar
  • 11 Elzagallaai A, Rose SD, Brandan NC. et al. Myristoylated alanine-rich C kinase substrate phosphorylation is involved in thrombin-induced serotonin release from platelets. Br J Haematol 2001; 112: 593-602.
    CrossrefPubMedGoogle Scholar
  • 12 Wood EG, Gray BM. Type-specific antibody prevents platelet aggregation induced by group B streptococci type III. J Lab Clin Med 1986; 107: 322-6.
    PubMedGoogle Scholar
  • 13 Peevy KJ, Panus P, Longenecker GL. et al. Prostaglandin synthetase inhibition in group B streptococcal shock: hematologic and hemodynamic effects. Pediatr Res 1986; 20: 864-6.
    CrossrefPubMedGoogle Scholar
  • 14 Pietrocola G, Schubert A, Visai L. et al. FbsA, a fibrinogen-binding protein from Streptococcus agalactiae, mediates platelet aggregation. Blood 2005; 105: 1052-9.
    CrossrefPubMedGoogle Scholar
  • 15 Jonsson IM, Pietrocola G, Speziale P. et al. Role of fibrinogen-binding adhesin expression in septic arthritis and septicemia caused by Streptococcus agalactiae . J Infect Dis 2005; 192: 1456-64.
    CrossrefPubMedGoogle Scholar
  • 16 Yeaman MR, Norman DC, Bayer AS. Staphylococcus aureus susceptibility to thrombin-induced platelet microbicidal protein is independent of platelet adherence and aggregation in vitro . Infect Immun 1992; 60: 2368-74.
    PubMedGoogle Scholar
  • 17 Ford I, Douglas CW, Cox D. et al. The role of immunoglobulin G and fibrinogen in platelet aggregation by Streptococcus sanguis . Br J Haematol 1997; 97: 737-46.
    CrossrefPubMedGoogle Scholar
  • 18 Pampolina C, McNicol A. Streptococcus sanguis-induced platelet activation involves two waves of tyrosine phosphorylation mediated by FcγRIIA and αIIbβ3. Thromb Haemost 2005; 93: 932-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Berner R, Bender A, Rensing C. et al. Low prevalence of the immunoglobulin-A-binding beta antigen of the C protein among Streptococcus agalactiae isolates causing neonatal sepsis. Eur J Clin Microbiol Infect Dis 1999; 18: 545-50.
    CrossrefPubMedGoogle Scholar
  • 20 Geiger J, Honig-Liedl P, Schanzenbacher P. et al. Ligand specificity and ticlopidine effects distinguish three human platelet ADP receptors. Eur J Pharmacol 1998; 351: 235-46.
    CrossrefPubMedGoogle Scholar
  • 21 Schwarz UR, Geiger J, Walter U. et al. Flow cytometry analysis of intracellular VASP phosphorylation for the assessment of activating and inhibitory signal transduction pathways in human platelets – definition and detection of ticlopidine/clopidogrel effects. Thromb Haemost 1999; 82: 1145-52.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Ledergerber D, Hartmann RW. Development of a screening assay for the in vitro evaluation of thromboxane A2 synthase inhibitors. J Enzyme Inhib 1995; 09: 253-61.
    PubMedGoogle Scholar
  • 23 Elzagallaai A, Rose SD, Trifaro JM. Platelet secretion induced by phorbol esters stimulation is mediated through phosphorylation of MARCKS: a MARCKSderived peptide blocks MARCKS phosphorylation and serotonin release without affecting pleckstrin phosphorylation. Blood 2000; 95: 894-902.
    PubMedGoogle Scholar
  • 24 Vallejo JG, Knuefermann P, Mann DL. et al. Group B streptococcus induces TNF-alpha gene expression and activation of the transcription factors NF-kappa B and activator protein-1 in human cord blood monocytes. J Immunol 2000; 165: 419-25.
    CrossrefPubMedGoogle Scholar
  • 25 Mancuso G, Midiri A, Beninati C. et al. Mitogen-activated protein kinases and NF-kappaB are involved in TNF-alpha responses to group B streptococci. J Immunol 2002; 169: 1401-9.
    CrossrefPubMedGoogle Scholar
  • 26 Fettucciari K, Fetriconi I, Bartoli A. et al. Involvement of mitogen-activated protein kinases in group B streptococcus-induced macrophage apoptosis. Pharmacol Res 2003; 47: 355-62.
    CrossrefPubMedGoogle Scholar
  • 27 Achison M, Elton CM, Hargreaves PG. et al. Integrin-independent tyrosine phosphorylation of p125(fak) in human platelets stimulated by collagen. J Biol Chem 2001; 276: 3167-74.
    CrossrefPubMedGoogle Scholar
  • 28 Wang XF, Liu F, Morris SA. et al. Synergistic outside-in regulation of platelet activation by GPIIb/IIIa ligand-induced conformation and oligomerization. Thromb Res 2001; 104: 239-47.
    CrossrefPubMedGoogle Scholar
  • 29 Canobbio I, Lova P, Sinigaglia F. et al. Proline-rich tyrosine kinase 2 and focal adhesion kinase are involved in different phases of platelet activation by vWF. Thromb Haemost 2002; 87: 509-17.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Mekrache M, Bachelot-Loza C, Ajzenberg N. et al. Activation of pp125FAK by type 2B recombinant von Willebrand factor binding to platelet GPIb at a high shear rate occurs independently of alpha IIb beta3 engagement. Blood 2003; 101: 4363-71.
    CrossrefPubMedGoogle Scholar
  • 31 Iwaki M, Kamachi K, Heveker N. et al. Suppression of platelet aggregation by Bordetella pertussis adenylate cyclase toxin. Infect Immun 1999; 67: 2763-8.
    PubMedGoogle Scholar
  • 32 Sheu JR, Hung WC, Kan YC. et al. Mechanisms involved in the antiplatelet activity of Escherichia coli lipopolysaccharide in human platelets. Br J Haematol 1998; 103: 29-38.
    CrossrefPubMedGoogle Scholar
  • 33 Sheu JR, Hung WC, Su CH. et al. The antiplatelet activity of Escherichia coli lipopolysaccharide is mediated through a nitric oxide/cyclic GMP pathway. Eur J Haematol 1999; 62: 317-26.
    PubMedGoogle Scholar
  • 34 Cherkasova TD, Bochkov VN, Iurkiv VA. et al. The capacity of Yersinia pestis toxin to decrease the functional response of human thrombocytes to stimulation by hormones. Biull Eksp Biol Med 1993; 116: 529-32.
    PubMedGoogle Scholar
  • 35 Cherkasova TD, Iurkiv VA. Features of regulatory system function in platelets under the effect of plague toxin. Vopr Med Khim 1994; 40: 17-20.
    PubMedGoogle Scholar
  • 36 Kalvegren H, Majeed M, Bengtsson T. Chlamydia pneumoniae binds to platelets and triggers P-selectin expression and aggregation: a causal role in cardiovascular disease?. Arterioscler Thromb Vasc Biol 2003; 23: 1677-83.
    CrossrefPubMedGoogle Scholar
  • 37 Bryant AE, Chen RY, Nagata Y. et al. Clostridial gas gangrene. II. Phospholipase C-induced activation of platelet gpIIbIIIa mediates vascular occlusion and myonecrosis in Clostridium perfringens gas gangrene. J Infect Dis 2000; 182: 808-15.
    CrossrefPubMedGoogle Scholar
  • 38 Flores-Diaz M, Alape-Giron A. Role of Clostridium perfringens phospholipase C in the pathogenesis of gas gangrene. Toxicon 2003; 42: 979-86.
    CrossrefPubMedGoogle Scholar
  • 39 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: 1033-44.
    CrossrefPubMedGoogle Scholar
  • 40 Sjobring U, Ringdahl U, Ruggeri ZM. Induction of platelet thrombi by bacteria and antibodies. Blood 2002; 100: 4470-7.
    CrossrefPubMedGoogle Scholar
  • 41 Kerrigan SW, Douglas I, Wray A. et al. A role for glycoprotein Ib in Streptococcus sanguis-induced platelet aggregation. Blood 2002; 100: 509-16.
    CrossrefPubMedGoogle Scholar
  • 42 Sullam PM, Jarvis GA, Valone FH. Role of immunoglobulin G in platelet aggregation by viridans group streptococci. Infect Immun 1988; 56: 2907-11.
    PubMedGoogle Scholar
  • 43 Deckmyn H, Vanhoorelbeke K, Peerlinck K. Inhibitory and activating human antiplatelet antibodies. Baillieres Clin Haematol 1998; 11: 343-59.
    CrossrefPubMedGoogle Scholar
  • 44 Brodeur BR, Boyer M, Charlebois I. et al. Identification of group B streptococcal Sip protein, which elicits cross-protective immunity. Infect Immun 2000; 68: 5610-8.
    CrossrefPubMedGoogle Scholar
  • 45 Holmsen H. Physiological functions of platelets. Ann Med 1989; 21: 23-30.
    CrossrefPubMedGoogle Scholar
  • 46 Tabuchi A, Yoshioka A, Higashi T. et al. Direct demonstration of involvement of protein kinase Calpha in the Ca2+-induced platelet aggregation. J Biol Chem 2003; 278: 26374-9.
    CrossrefPubMedGoogle Scholar
  • 47 Ozaki Y, Jinnai Y, Yatomi Y. et al. Protein kinase C inhibitors suppress disc-sphere changes of human platelets, as assessed with the shape-change parameter. Eur J Pharmacol 1993; 235: 255-65.
    CrossrefPubMedGoogle Scholar
  • 48 Murphy CT, Westwick J. Selective inhibition of protein kinase C. Effect on platelet-activating-factorinduced platelet functional responses. Biochem J 1992; 283: 159-64.
    CrossrefPubMedGoogle Scholar
  • 49 Murugappan S, Tuluc F, Dorsam RT. et al. Differential role of protein kinase C delta isoform in agonist-induced dense granule secretion in human platelets. J Biol Chem 2004; 279: 2360-7.
    CrossrefPubMedGoogle Scholar
  • 50 Rosado JA, Sage SO. Role of the ERK pathway in the activation of store-mediated calcium entry in human platelets. J Biol Chem 2001; 276: 15659-65.
    CrossrefPubMedGoogle Scholar
  • 51 McNicol A, Jackson EC. Inhibition of the MEK/ ERK pathway has no effect on agonist-induced aggregation of human platelets. Biochem Pharmacol 2003; 65: 1243-50.
    CrossrefPubMedGoogle Scholar
  • 52 Kramer RM, Roberts EF, Strifler BA. et al. Thrombin induces activation of p38 MAP kinase in human platelets. J Biol Chem 1995; 270: 27395-8.
    CrossrefPubMedGoogle Scholar
  • 53 Saklatvala J, Rawlinson L, Waller RJ. et al. Role for p38 mitogen-activated protein kinase in platelet aggregation caused by collagen or a thromboxane analogue. J Biol Chem 1996; 271: 6586-9.
    CrossrefPubMedGoogle Scholar
  • 54 Bugaud F, Nadal-Wollbold F, Levy-Toledano S. et al. Regulation of c-jun-NH2 terminal kinase and extracellular-signal regulated kinase in human platelets. Blood 1999; 94: 3800-5.
    PubMedGoogle Scholar
  • 55 Schwarz UR, Kobsar AL, Koksch M. et al. Inhibition of agonist-induced p42 and p38 mitogen-activated protein kinase phosphorylation and CD40 ligand/P-selectin expression by cyclic nucleotide-regulated pathways in human platelets. Biochem Pharmacol 2000; 60: 1399-407.
    CrossrefPubMedGoogle Scholar
  • 56 Hackeng CM, Relou IA, Pladet MW. et al. Early platelet activation by low density lipoprotein via p38MAP kinase. Thromb Haemost 1999; 82: 1749-56.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 57 Canobbio I, Reineri S, Sinigaglia F. et al. A role for p38 MAP kinase in platelet activation by von Willebrand factor. Thromb Haemost 2004; 91: 102-10.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 58 Dash D, Aepfelbacher M, Siess W. The association of pp125FAK, pp60Src, CDC42Hs and Rap1B with the cytoskeleton of aggregated platelets is a reversible process regulated by calcium. FEBS Lett 1995; 363: 231-4.
    CrossrefPubMedGoogle Scholar
  • 59 Lyman S, Gilmore A, Burridge K. et al. Integrinmediated activation of focal adhesion kinase is independent of focal adhesion formation or integrin activation. Studies with activated and inhibitory beta3 cytoplasmic domain mutants. J Biol Chem 1997; 272: 22538-47.
    CrossrefPubMedGoogle Scholar