Molecular and Cellular Biochemistry

, Volume 288, Issue 1–2, pp 171–178 | Cite as

Staphylococcal Enterotoxin B Initiates Protein Kinase C Translocation and Eicosanoid Metabolism While Inhibiting Thrombin-Induced Aggregation in Human Platelets

  • Uyen Tran
  • Thomas Boyle
  • Jeffrey W. Shupp
  • Rasha Hammamieh
  • Marti Jett


Staphylococcal enterotoxin (SE) B, a heat-stable toxin secreted by Staphylococcus aureus, has been implicated in the pathogenesis and exacerbation of several critical illnesses. It has been hypothesized that enterotoxins may interact with blood products such as platelets, in addition to T-lymphocytes and renal proximal tubule cells. The aim of this present study was to elucidate whether SEB directly alters human platelet function. Human platelet rich plasma (PRP) was pre-incubated with SEA, SEB, SEC or TSST-1, (at various concentrations and incubation times). After incubation, PRP was exposed to thrombin and aggregation was assessed. Incubation with all toxins tested resulted in decreased aggregation, specifically; exposure to 10μ g/ml of SEB for 30 min caused a 20% decrease and a 49% decrease at 90 min. A similar reduction in aggregation was seen in samples incubated with phorbol myristate acetate, a known stimulator of protein kinase C (PKC). Further, platelets exposed to SEB exhibited an increased plasma membrane PKC activity. Sphingosine, an inhibitor of PKC proved to block the SEB-induced reduction in aggregation. SEB effects on platelet metabolism were investigated using high performance liquid chromatography showing up to a 2-fold increase of active metabolites lipoxin A4 and 12-HETE, as compared to control. These data indicate that SEB is able to induce platelet dysfunction, and these effects may be mediated through activation of PKC.


aggregation platelets SEB shock superantigen 


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  1. 1.
    Al-Sarraf AA, Christenson JT, Owunwanne A: Early and late platelet sequestration in different organs during endotoxic shock. Res Exp Med (Berl) 188 (1): 59–66, 1988CrossRefGoogle Scholar
  2. 2.
    Barry BE, Crapo JD: Patterns of accumulation of platelets and neutrophils in rat lungs during exposure to 100% and 85% oxygen. Am Rev Respir Dis 132(3): 548–555, 1985PubMedGoogle Scholar
  3. 3.
    Bhatia M, Moochhala S: Role of inflammatory mediators in the pathophysiology of acute respiratory distress syndrome. J Pathol 202(2): 145–156, 2004PubMedCrossRefGoogle Scholar
  4. 4.
    Boogaerts MA, Yamada O, Jacob HS, Moldow CF: Enhancement of granulocyte-endothelial cell adherence and granulocyte-induced cytotoxicity by platelet release products. Proc Natl Acad Sci USA 79(22): 7019–7023, 1982PubMedCrossRefADSGoogle Scholar
  5. 5.
    Bowers RE, Ellis EF, Brigham KL, Oates JA: Effects of prostaglandin cyclic endoperoxides on the lung circulation of unanesthetized sheep. J Clin Invest 63(1): 131–137, 1979PubMedGoogle Scholar
  6. 6.
    Boyle T, Lancaster V, Hunt R, Gemski P, Jett M: Method for simultaneous isolation and quantitation of platelet activating factor and multiple arachidonate metabolites from small samples: analysis of effects of Staphylococcus aureus enterotoxin B in mice. Anal Biochem 216(2): 373–382, 1994PubMedCrossRefGoogle Scholar
  7. 7.
    Bull BS, Bull MH: Hypothesis: disseminated intravascular inflammation as the inflammatory counterpart to disseminated intravascular coagulation. Proc Natl Acad Sci USA 91(17): 8190–8194, 1994PubMedCrossRefADSGoogle Scholar
  8. 8.
    Campbell WN, Fitzpatrick M, Ding X, Jett M, Gemski P, Goldblum SE: SEB is cytotoxic and alters EC barrier function through protein tyrosine phosphorylation in vitro. Am J Physiol 273(1 Pt 1): L31-39, 1997PubMedGoogle Scholar
  9. 9.
    Chatterjee S, Jett M: Glycosphingolipids: the putative receptor for Staphylococcus aureus enterotoxin-B in human kidney proximal tubular cells. Mol Cell Biochem 113(1): 25–31, 1992PubMedCrossRefGoogle Scholar
  10. 10.
    Fratantoni JC, Poindexter BJ: Measuring platelet aggregation with microplate reader. A new technical approach to platelet aggregation studies. Am J Clin Pathol 94(5): 613–617, 1990Google Scholar
  11. 11.
    Gareau R, Gruda J, Micusan VV: Effect of toxic shock syndrome toxin-1 on human hemostatic parameters. Thromb Res 54(4): 349–356, 1989PubMedCrossRefGoogle Scholar
  12. 12.
    Goetzl EJ, Valone FH, Reinhold VN, Gorman RR: Specific inhibition of the polymorphonuclear leukocyte chemotactic response to hydroxy-fatty acid metabolites of arachidonic acid by methyl ester derivatives. J Clin Invest 63(6): 1181–1186, 1979PubMedGoogle Scholar
  13. 13.
    Grabarek J, Timmons S, Hawiger J: Modulation of human platelet protein kinase C by endotoxic lipid A. J Clin Invest 82(3): 964–971, 1988PubMedGoogle Scholar
  14. 14.
    Hannun YA, Greenberg CS, Bell RM: Sphingosine inhibition of agonist-dependent secretion and activation of human platelets implies that protein kinase C is a necessary and common event of the signal transduction pathways. J Biol Chem 262(28): 13620–13626, 1987PubMedGoogle Scholar
  15. 15.
    Henghold WB: 2nd. Other biologic toxin bioweapons: ricin, staphylococcal enterotoxin B, and trichothecene mycotoxins. Dermatol Clin 22(3): 257–262, v, 2004PubMedCrossRefGoogle Scholar
  16. 16.
    Idell S: Coagulation, fibrinolysis, and fibrin deposition in acute lung injury. Crit Care Med 31 (4 Suppl): S213–220, 2003PubMedCrossRefGoogle Scholar
  17. 17.
    Iwaki M, Kamachi K, Heveker N, Konda T: Suppression of platelet aggregation by Bordetella pertussis adenylate cyclase toxin. Infect Immun 67(6): 2763–2768, 1999PubMedGoogle Scholar
  18. 18.
    Jett M, Brinkley W, Neill R, Gemski P, Hunt R: Staphylococcus aureus enterotoxin B challenge of monkeys: correlation of plasma levels of arachidonic acid cascade products with occurrence of illness. Infect Immun 58(11): 3494–3499, 1990PubMedGoogle Scholar
  19. 19.
    Jett M, Ionin B, Das R Neill R: The Staphylococcal Enterotoxins. In: Sussman M, ed. Molecular Medical Microbiology. San Diego: Academic Press, 2002Google Scholar
  20. 20.
    Marcus AJ, Broekman MJ, Safier LB, Ullman HL, Islam N, Sherhan CN, Rutherford LE, Korchak HM, Weissmann G: Formation of leukotrienes and other hydroxy acids during platelet-neutrophil interactions in vitro. Biochem Biophys Res Commun 109(1): 130–137, 1982PubMedCrossRefGoogle Scholar
  21. 21.
    Nishioka H, Horiuchi H, Tabuchi A, Yoshioka A, Shirakawa R, Kita T: Small GTPase Rho regulates thrombin-induced platelet aggregation. Biochem Biophys Res Commun 280(4): 970–975, 2001PubMedCrossRefGoogle Scholar
  22. 22.
    Schantz EJ, Roessler WG, Wagman J, Spero L, Dunnery DA, Bergdoll MS: Purification of staphylococcal enterotoxin B. Biochemistry 4(6): 1011–1016, 1965PubMedCrossRefGoogle Scholar
  23. 23.
    Sheagren JN: Staphylococcal infections of the skin and skin structures. Cutis 36(5A): 2–6, 1985PubMedGoogle Scholar
  24. 24.
    Shupp JW, Jett M, Pontzer CH: Identification of a transcytosis epitope on staphylococcal enterotoxins. Infect Immun 70(4): 2178–2186, 2002PubMedCrossRefGoogle Scholar
  25. 25.
    Sigurdsson GH, Christenson JT, el-Rakshy MB, Sadek S: Intestinal platelet trapping after traumatic and septic shock. An early sign of sepsis and multiorgan failure in critically ill patients? Crit Care Med 20(4): 458–467, 1992PubMedGoogle Scholar
  26. 26.
    Sun G, Chang WL, Li J, Berney SM, Kimpel D, van der Heyde HC: Inhibition of platelet adherence to brain microvasculature protects against severe Plasmodium berghei malaria. Infect Immun 71(11): 6553–6561, 2003PubMedCrossRefGoogle Scholar
  27. 27.
    Tabuchi A, Yoshioka A, Higashi T, Shirakawa R, Nishioka H, Kita T, Horiuchi H: Direct demonstration of involvement of protein kinase Calpha in the Ca2+-induced platelet aggregation. J Biol Chem 278(29): 26374–26379, 2003PubMedCrossRefGoogle Scholar
  28. 28.
    Utsonomiya T, Krausz MM, Levine L, Shepro D, Hechtman HB: Thromboxane mediation of cardiopulmonary effects of embolism. J Clin Invest 70(2): 361–368, 1982PubMedCrossRefGoogle Scholar
  29. 29.
    van Gessel YA, Mani S, Bi S, Hammamieh R, Shupp JW, Das R, Coleman GD, Jett M: Functional piglet model for the clinical syndrome and postmortem findings induced by staphylococcal enterotoxin B. Exp Biol Med (Maywood) 229(10): 1061–1071, 2004Google Scholar
  30. 30.
    Villavicencio RT, Wall MJ, Jr: The pathogenesis of Staphylococcus aureus in the trauma patient and potential future therapies. Am J Surg 172(3): 291–296, 1996PubMedCrossRefGoogle Scholar
  31. 31.
    Yan Z, Yang DC, Neill R, Jett M: Production of tumor necrosis factor alpha in human T lymphocytes by staphylococcal enterotoxin B correlates with toxin-induced proliferation and is regulated through protein kinase C. Infect Immun 67(12): 6611–6618, 1999PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Uyen Tran
    • 1
    • 2
  • Thomas Boyle
    • 1
    • 3
  • Jeffrey W. Shupp
    • 1
  • Rasha Hammamieh
    • 1
  • Marti Jett
    • 1
  1. 1.Department of Molecular PathologyWalter Reed Army Institute of ResearchSilver SpringUSA
  2. 2.Vanderbilt University Medical CenterNashvilleUSA
  3. 3.FDA/CFSANCollege ParkUSA

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