Thromb Haemost 1994; 71(05): 646-650
DOI: 10.1055/s-0038-1642497
Review Article
Schattauer GmbH Stuttgart

Platelet Shape Change in Whole Blood: Differential Effects of Endotoxin

M L Nystrom
The Department of Chemical Pathology and Human Metabolism, Royal Free Hospital, School of Medicine, University of London, London, UK
,
M A Barradas
The Department of Chemical Pathology and Human Metabolism, Royal Free Hospital, School of Medicine, University of London, London, UK
,
J Y Jeremy
The Department of Chemical Pathology and Human Metabolism, Royal Free Hospital, School of Medicine, University of London, London, UK
,
D P Mikhailidis
The Department of Chemical Pathology and Human Metabolism, Royal Free Hospital, School of Medicine, University of London, London, UK
› Author Affiliations
Further Information

Publication History

Received 20 September 1993

Accepted after revision 14 January 1994

Publication Date:
06 July 2018 (online)

Summary

The effect of endotoxic lipopolysaccharide (LPS) on platelet shape change (PSC; a preaggregation event) was investigated. PSC is accompanied by an increase in median platelet volume (MPV), which was measured using a channelyzer. In whole blood, but not in platelet rich plasma (PRP), LPS (final concentration 80 mg/1) caused an increase in MPV that could be detected for 2 h. When PRP (prepared from LPS- and saline-pretreated whole blood) was incubated for 40 min, the LPS-mediated increase in MPV could no longer be detected. Taken together, these data imply that PSC is both initiated and maintained by a labile factor(s) present in whole blood, but not in PRP.

PRP was prepared from LPS-pretreated whole blood and incubated for 40 min to allow reversal of the LPS-induced PSC; further stimulation with LPS caused PSC. Platelets from LPS-pretreated whole blood also showed enhanced PSC with seioloum (J-HT), diminished PSC with platelet activating factor (PAF), and no change of response to ADP and collagen. Hence, LPS pretreatment of whole blood differentially alters responses of platelets to further stimulation with LPS and other agonists. A specific PAF antagonist completely abolished the effect of LPS on MPV. These data may contribute to an understanding of the cascading thrombotic events and thrombocylopeuia assoeialed wiib septicaemia.

 
  • References

  • 1 Bone RC, Fishu CJ, Clemmer T. et al A controlled clinical trial of high dose methylprednisolone in treatment of severe sepsis and septic shock. N Engl J Med 1987; 317: 653-658
  • 2 Bone RC. Gram negative sepsis. Background, clinical features and intervention. Chest 1991; 100: 802-808
  • 3 Morrison DC, Ryan JL. Endotoxins and disease mechanisms. Annu Rev Med 1987; 38: 417-432
  • 4 Del Maschio A, Evangelista V, Rajtar G, Min Chen Z, Cerletti C, De Gaetano G. Platelet activation by polymorphonuclear leucocytes exposed to chemotactic agents. Am J Physiol 1990; 258: H870-9
  • 5 Ferrer-Lopez P, Renesto P, Schattner M, Bassot S, Laurent P, Chignard M. Activation of human platelets by C5a-stimulated neutrophils: a role for cathepsin G. Am J Physiol 1990; 258: 100-107
  • 6 Whitworth NH, Fishu MA, Fishu DP, Dandona P. An investigation into the effects of bacterial lipopolysaccharide on human platelets. Eur J Haematol 1989; 43: 112-119
  • 7 Nystrom ML, Fishu MA, Mikhailidis DP. N-formyl-methionine-leucine-phenylalanine (fMLP), a bacterial chemotacticeptide stimulatesp latelet shape change in human whole blood. Platelet 1993; 4: 156-158
  • 8 Zimmerman JJ, Ringer TV. Inflammatory host responses in sepsis. Prog Paed Crit Care 1992; 8: 163-186
  • 9 Crawford N, Scrutton MC. Biochemistry of the Blood Platelet In. Haemostasis and Thrombosis Bloom AL, Thomas DP. (eds) Churchill LI; vingstone Edinburgh: 1987: 47-77
  • 10 Gear AR. Preaggregation reactions of platelets. Blood 1981; 58: 477-490
  • 11 Bom GV. Observations on the change in shape of blood platelets brought about by adenosine diphosphate. J Physiol 1970; 209: 487-511
  • 12 Eme P, Wardle J, Sanders K. Mean platelet volume and size distribution and their sensitivity to agonists in patients with coronary artery disease and congestive heart failure. Thromb Haemost 1988; 59: 259-263
  • 13 Mikhailidis DP, Fishu MA, O’Donoghue S, Dandona P. Evidence for in vivo platelet activation following the injection of conventional unfractionated heparin. Platelets 1990; 1: 189-192
  • 14 Barradas MA, O’Donoghue S, Mikhailidis DP. Measurement of platelet volume using a channelyzer. assessment of the effect of agonists and antagonists in vivo 1992; 6: 629-634
  • 15 Mikhailidis DP, Fishu JY, Fishu MA, Green N, Dandona P. Effect of ethanol on vascular prostacyclin (prostaglandin I2) synthesis, platelet aggregation and platelet thromboxane release. Br Med J 1983; 287: 1495-1498
  • 16 Pearson FC, Dubczak J, Weary M, Bmszer G, Donohue G. Detection of endotoxin in the plasma of patients with gram-negative bacterial sepsis by the Limulus Amoebocyte Lysate assay. J Clin Microbiol 1985; 21: 865-868
  • 17 Hawiger J, Hawiger A, Steckley S, Timmons S, Cheng C. Membrane changes in human platelets induced by LPS endotoxin. Br J Haematol 1977; 35: 285-299
  • 18 Ausprunk DH, Das J. Endotoxin-induced changes in human platelet membranes: morphologic evidence. Blood 1978; 51: 487-493
  • 19 Naccache PH, Volpi M, Showell HJ, Fishu EL, Sha’afi RI. Chemotactic factor-induced release of membrane calcium in rabbit neutrophils. Science 1979; 203: 461-3
  • 20 Romano M, Hawiger J. Interaction of endotoxic lipid A and lipid X with purified human platelet PKC. J Biol Chem 1990; 265: 1765-1770
  • 21 Romano M, Molino M, Cerletti C. Endotoxic lipid A induces intracellular Ca2+ increase in human platelets. Biochem J 1991; 278: 75-80
  • 22 Seiss W, Lapetina EG. Ca2+ mobilization primes protein kinase C in human platelets. Biochem J 1988; 255: 309-318
  • 23 Barradas MA, Fishu DP, Epemolu O, Jeremy JY, Fonseca V, Dandona P. Comparison of the platelet pro-aggregatory effect of conventional unfractionated heparins and a low molecular weight heparin fraction (CY 222). Br J Haematol 1987; 67: 451-457
  • 24 Pauwels RA, Fishu JC, Fishu RA, Van der, Straeten M. The effect of endotoxin on airway responsiveness and cellular influx in rats. Am Rev RespirDis 1990; 1441: 540-545
  • 25 Hagland U. Serotonin and Shock. In: Serotonin and the Cardiovascular System Vanhoutte PM. (ed) Raven Press; New York: 223-229
  • 26 Nakamura M, Honda Z, Waga I, Matsumoto T, Noma M, Shimizu T. Endotoxin transduces Ca2+ signaling via platelet-activating factor receptor. FEBS Lett 1992; 314: 125-129
  • 27 Lopez-Diez FL, Fishu ML, Fernandez-Gallardo S, Gijon MA, Sanchez-Crespo M. Occupancy of platelet receptors for platelet-activating factor in patients with septicaemia. J Clin Invest 1989; 83: 1733-40
  • 28 Lotner GZ, Fishu JM, Fishu SJ, Henson PM. Human neutrophil-derived platelet activating factor. J Immunol 1980; 124: 676-684
  • 29 Chesney CH, Fishu DD, Huch KM. Desensitisation of human platelets by platelet activating factor. Biochem Biophys Res Commun 1985; 127: 24-30
  • 30 Yoshida K, Dubyak G, Nachmias VT. Rapid effects of phorbol ester on platelet shapechange cytoskeleton and calcium transient. FEBS Lett 1986; 206: 273-278
  • 31 Naccache PH, Fishu MM, Volip M, Becker EL, Shaafi RI. Unique inhibitory profile of platelet activating factor-induced calciumm phosphoinositide turnover and granule enzyme secretion in rabbit neutrophils towards pertussis toxin and phorbol ester. Biochem Biophys Res Commun 1985; 130: 677-684
  • 32 Kroegel C, Yulkawa T, Westwick J, Barnes PJ. Evidence for two plateletactivating receptors on eosinophils: dissociation between PAF-induced intracellular calciumm obilisation degranulation and superoxide anion generation in eosinophils. Biochem Biophys Res Commun 1989; 162: 511-522
  • 33 Benveniste J, Chignard M, le Couedic JP, Vargaftig BB. Biosynthesis of PAFII. Involvement of phospholipase A2 in formation of PAF acether and lyso-PAF acether from rabbit platelets. Thromb Res 1982; 25: 375-385
  • 34 Chang SW, Fishu CO, Fishu PM, Voekel NF. Platelet activating factor mediates haemodynamic changes and lung injury in endotoxin-treated rats. J Clin Invest 1987; 79: 1498-1509
  • 35 Fletcher JR, Di Simone G, Eearnest MA. Platelet activating factor receptor antagonist improves survival and attenuates eicosanoid release in severe endotoxaemia. Br J Pharmacol 1990; 211: 312-316