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Thromb Haemost 1988; 59(03): 378-382
DOI: 10.1055/s-0038-1647499
Original Article
Schattauer GmbH Stuttgart

Endotoxin-Induced Platelet Activation in Human Whole Blood In Vitro

Gyorgy Csako
1   The Clinical Pathology Department, Clinical Center, National lnstitutes of Health, Bethesda, MD, USA
,
Eva A Suba
2   The Naval Medical Research Institute, Bethesda, MD, USA
,
Ronald J Elin
1   The Clinical Pathology Department, Clinical Center, National lnstitutes of Health, Bethesda, MD, USA
› Institutsangaben
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Publikationsverlauf

Received 27. Juli 1987

Accepted after revision 11. Januar 1988

Publikationsdatum:
29. Juni 2018 (online)

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Summary

The effect of purified bacterial endotoxin was studied on human platelets in vitro. In adding up to 1 μg/mL of a highly purified endotoxin, we found neither aggregation nor ATP release in heparinized or citrated human platelet-rich plasma. On the other hand, endotoxin at concentrations as low as a few ng/mL (as may be found in septic patients) caused platelet aggregation in both heparinized and citrated human whole blood, as monitored by change in impedance, free platelet count, and size. Unlike collagen, the platelet aggregation with endotoxin occurred after a long lag phase, developed slowly, and was rarely coupled with measurable release of ATP. The platelet aggregating effect of endotoxin was dose-dependent and modified by exposure of the endotoxin to ionizing radiation. Thus, the activation of human platelets by “solubilized” endotoxin in plasma requires the presence of other blood cells. We propose that the platelet effect is mediated by monocytes and/or neutrophils stimulated by endotoxin.

Keywords

Platelet aggregation - ATP release - Whole blood - Bacterial lipopolysaccharide - Ionizing radiation
 

  • References

  • 1 Stanford JP. Epidemiology and overview of the problem. In: Septic shock. Root RK, Sande MA. (eds) Churchill Livingstone; New York: 1985: 01-11
    Google Scholar
  • 2 Luderitz O, Galanos C, Rietschel ET. Endotoxins of gram-negative bacteria. In: International encyclopedia of pharmacology and therapeutics. Section 119. Pharmacology of bacterial toxins. Dorner F, Drews J. Pergamon Press; Oxford: 1986: 307-326
    Google Scholar
  • 3 Wilson JJ, Neame PB, Kelton JG. Infection-induced thrombocy-topenia. Semin Thromb Haemost 1982; 08: 217-233
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 4 Myrvold HE, Brandburg A, Lindquist O, Busch C, Lewis DH. The role of platelets and fibrinogen in experimental septic shock. Acta Chir Scand 1977; 143: 131-137
    PubMedGoogle Scholar
  • 5 Gralnick HR, Elin R, Abrans E, Givelber H, Wolff SM. Effects of endotoxin (LPS) on blood coagulation in humans. Clin Res 1976; 24: 344A
    PubMedGoogle Scholar
  • 6 Cohen P, Braunwald J, Gardner FH. Destruction of canine platelets and rabbit platelets following intravenous administration of carbon particles or endotoxin. J Lab Clin Med 1965; 66: 263-271
    PubMedGoogle Scholar
  • 7 Fletcher JG, Ramwell PW. The effects of prostacyclin (PGI2) on endotoxin shock and endotoxin-induced platelet aggregation in dogs. Circ Shock 1980; 07: 299-308
    PubMedGoogle Scholar
  • 8 Spielvogel AR. An ultrastructural study of the mechanisms of platelet-endotoxin interaction. J Exp Med 1967; 126: 235-250
    CrossrefPubMedGoogle Scholar
  • 9 Gröttum KA, Hjort PF, Jeremic M. Interaction of bacterial endotoxin and liquoid with blood platelets: Aggregation and decrease in the electrokinetic charge. Thromb Diath Haemorrh 1969; 22: 192-202
    PubMedGoogle Scholar
  • 10 Lewis AF, Dickson RC. Platelet aggregation: Induction by means of a complex between endotoxin and copper (Cu2+). Can J Biochem 1971; 49: 1236-1244
    CrossrefPubMedGoogle Scholar
  • 11 Nagayama M, Zucker MB, Beller FK. Effects of a variety of endotoxins on human and rabbit platelet function. Thromb Diath Haemorrh 1971; 26: 467-473
    PubMedGoogle Scholar
  • 12 MacIntyre DE, Allen AP, Thome KJI, Glautert AM, Gordon JL. Endotoxin-induced platelet aggregation and secretion. Morphological changes and pharmacological effects. J Cell Sci 1977; 28: 211-223
    PubMedGoogle Scholar
  • 13 Abdelnoor AM, Kassem H, Bikhazi AB, Nowotny A. Effect of gram-negative bacterial lipopolysaccharide-derived polysaccharides, glycolipids, and lipopolysaccharides on rabbit and human platelets in vitro. Immunobiol 1980; 157: 145-153
    CrossrefPubMedGoogle Scholar
  • 14 Morrison DC, Oades ZG, DePietro D. Endotoxin-initiated membrane changes in rabbit platelets. Progr Clin Biol Res 1981; 62: 47-64
    PubMedGoogle Scholar
  • 15 Mueller-Eckhardt C, Luscher EF. Studies of the mechanisms of the human platelet release reaction induced by immunologic stimuli. II. The effect of zymosan. J Immunol 1974; 112: 1211-1218
    PubMedGoogle Scholar
  • 16 Csako G, Suba EA. A physiologic regulator of collagen-induced platelet aggregation: Inhibition by C1q. Thromb Haemostas 1981; 45: 110-115
    PubMedGoogle Scholar
  • 17 Ream VJ, Deykin D, Gurewich V, Wessler S. The aggregation of human platelets by bacterial endotoxin. J Lab Clin Med 1965; 66: 245-252
    PubMedGoogle Scholar
  • 18 Hawiger J, Hawiger A, Steckley S, Timmons S, Cheng C. Membrane changes in human platelets induced by lipopolysaccharide endotoxin. Br J Haemat 1977; 35: 285-299
    CrossrefPubMedGoogle Scholar
  • 19 Ausprunk DH, Das J. Endotoxin-induced changes in human platelet membranes: Morphologic evidence. Blood 1978; 51: 487-495
    PubMedGoogle Scholar
  • 20 Ginsberg MH, Henson PM. Enhancement of platelet response to immune complexes and IgG aggregates by lipid A-rich bacterial lipopolysaccharides. J Exp Med 1978; 147: 207-218
    CrossrefPubMedGoogle Scholar
  • 21 Landes RR, Morrison DC. Biochemical mechanisms of lipid A-mediated enhancement of platelet secretory response to aggregated immunoglobulins. Am J Pathol 1981; 104: 196-205
    PubMedGoogle Scholar
  • 22 Stuart M. Effect of endotoxin on arachidonic acid release and thromboxane B2 production by human platelets. Am J Hemat 1981; 11: 159-164
    CrossrefPubMedGoogle Scholar
  • 23 Saba HI, Saba SR, Morelli G, Hartmann RC. Endotoxin-mediated inhibition of human platelet aggregation. Thromb Res 1984; 34: 19-33
    CrossrefPubMedGoogle Scholar
  • 24 Rudbach JA, Akiya FI, Elin RJ, Hochstein HD, Luoma MD, Milner ECB, Milner KC, Thomas KR. Preparation and properties of a national reference endotoxin. J Clin Microbiol 1976; 03: 21-25
    PubMedGoogle Scholar
  • 25 Csako G, Suba EA, Ahlgren A, Tsai C-M, Elin RJ. Relation of structure to function for the U.S. reference standard endotoxin after exposure to 60Co-radiation. J Infect Dis 1986; 153: 98-108
    CrossrefPubMedGoogle Scholar
  • 26 Abbate R, Favilla S, Boddi M, Costanzo G, Prisco D. Factors influencing platelet aggregation in whole blood. Am J Clin Pathol 1986; 86: 91-96
    CrossrefPubMedGoogle Scholar
  • 27 Mehta P, Mehta J, Ostrowski N, Aguila E. Potentiation of platelet aggregation by Chronolume. Thromb Res 1983; 32: 509-512
    CrossrefPubMedGoogle Scholar
  • 28 Timmons S, Huzoor Akbar, Grabarek J, Kloczewiak M, Hawiger J. Mechanism of human platelet activation by endotoxic glycolipid-bearing mutant Re595 of Salmonella minnesota. Blood 1986; 68: 1015-1023
    PubMedGoogle Scholar
  • 29 Chignard M, LeCouedic JP, Tence M, Vargaftig BB, Benveniste J. The role of platelet activating factor in platelet aggregation. Nature (Lond) 1979; 279: 799-800
    CrossrefPubMedGoogle Scholar
  • 30 Rivers RPA, Hathaway WE, Weston WL. The endotoxin-induced coagulant activity of human monocytes. Br J Haematol 1975; 30: 311-316
    CrossrefPubMedGoogle Scholar
  • 31 Schwartz BS, Monroe MC. Human platelet aggregation is initiated by peripheral blood mononuclear cells exposed to bacterial lipopolysaccharide in vitro. J Clin Invest 1986; 78: 1136-1141
    CrossrefPubMedGoogle Scholar
  • 32 Levin J, Poore TE, Zauber NP, Oser RS. Detection of endotoxin in the blood of patients with sepsis due to gram-negative bacteria. N Engl J Med 1970; 283: 1313-1316
    CrossrefPubMedGoogle Scholar
  • 33 Fink PC, Lehr L, Urbaschek RM, Kozak J. Limulus amebocyte lysate test for endotoxemia: Investigations with a femtogram sensitive spectrophotometric assay. Klin Wschr 1981; 59: 213-218
    CrossrefPubMedGoogle Scholar
  • 34 Elin RJ, Wolff SM, McAdam KPWJ, Chedid L, Audibert F, Bernard C, Oberling F. Properties of reference Escherichia coli endotoxin and its phthalylated derivative in humans. J Infect Dis 1981; 144: 329-336
    CrossrefPubMedGoogle Scholar
  • 35 Smedly LA, Tonnesen MG, Sandhaus RA, Haslett C, Guthrie LA, Johnston Jr RB, Henson PM, Worthen GS. Neutrophil-mediated injury to endothelial cells. Enhancement by endotoxin and essential role of neutrophil elastase J Clin Invest 1986; 77: 1233-1243
    CrossrefPubMedGoogle Scholar
  • 36 Mueller HW, O’Flahert JT, Wykle RL. Biosynthesis of platelet activating factor in rabbit polymorphonuclear neutrophils. J Biol Chem 1983; 258: 6213-6218
    PubMedGoogle Scholar
  • 37 Clark PO, Hanahan DJ, Pinckard RN. Physical and chemical properties of platelet-activating factor obtained from human neutrophils and monocytes and rabbit neutrophils and basophils. Biochim Biophys Acta 1980; 628: 69-75
    CrossrefPubMedGoogle Scholar
  • 38 Kuehl Jr FA, Egan RW. Prostaglandins, arachidonic acid, and inflammation. Science 1980; 210: 978-984
    CrossrefPubMedGoogle Scholar
  • 39 Semamaro N, Locati D, Colucci M. Leucocyte-dependent platelet activation: an alternative pathway for initiation of blood clotting in inflammation. Agents Actions 1981; 11: 553-555
    CrossrefPubMedGoogle Scholar
  • 40 Davis RB, Johnson MF. Effects of bacterial endotoxin and platelet activating facter (PAF) on human platelet aggregation in native whole blood. Thromb Res 1986; 44: 565-573
    CrossrefPubMedGoogle Scholar