Thromb Haemost 2000; 84(03): 381-387
DOI: 10.1055/s-0037-1614032
Commentary
Schattauer GmbH

Multi-Cellular Activation In Vivo by Endotoxin in Humans – Limited Protection by Adenosine Infusion

Nailin Li
1   From the Department of Medicine, Division of Clinical Pharmacology, Karolinska Hospital, Stockholm
,
Anne Soop
2   Division of Anesthesiology, Department of KARO, Huddinge Hospital, Huddinge, Sweden
,
Alf Sollevi
2   Division of Anesthesiology, Department of KARO, Huddinge Hospital, Huddinge, Sweden
,
Paul Hjemdahl
1   From the Department of Medicine, Division of Clinical Pharmacology, Karolinska Hospital, Stockholm
› Author Affiliations
The authors are grateful to Mrs Maud Daleskog, Pia Hillelson, Maj-Christina Johansson, Wiveca Gustavsson, and Inga Hellström for their expert technical assistance. The study was support by grants from the Swedish Medical Research Council (5930, 7485), the King Gustaf V and Queen Victoria Foundation, the Swedish Heart-Lung Foundation, the Swedish Society for Medical Research, and Item Development AB (Stockholm, Sweden).
Further Information

Publication History

Received 24 December 1999

Accepted after resubmission 04 April 2000

Publication Date:
14 December 2017 (online)

Summary

The influence of adenosine infusion (40 µg/kg/min for 4 h) on inflammatory and hemostatic parameters was investigated in healthy males without (n = 10) or with (n = 11) intravenous endotoxin injection (4 ng/kg). Without endotoxin, adenosine elevated circulating leukocytes and circulating platelet-leukocyte aggregates. Endotoxin activated platelets and leukocytes in vivo. Platelet activation was seen as slightly increased platelet P-selectin expression, decreased platelet counts, and elevated plasma soluble P-selectin (from 39.6 ± 3.4 to 68.9 ± 6.6 ng/ml; P <0.01). Leukocyte activation was evidenced by increased CD11b expression (from MFI of 0.54 ± 0.02 to 2.21 ± 0.17; P <0.01) and plasma elastase levels (from 25.3 ± 2.5 to 169.3 ± 22.5 ng/ml; P <0.01). Endotoxin also enhanced platelet and leukocyte responsiveness to in vitro stimulation. Endotoxin induced von Willebrand factor secretion (from 92 ± 8 units to 265 ± 19 units at 4 h; P <0.001) and enhanced thrombin generation in vivo. Endotoxin induced leukocytosis and thus increased circulating platelet-leukocyte, mainly platelet-neutrophil, aggregates. Adenosine caused slight attenuation of platelet reactivity to agonist stimulation, enhanced the endotoxin-induced leukocytosis, and detained more platelet-leukocyte aggregates in circulation, but did not attenuate endotoxin-induced neutrophil elastase secretion, von Willebrand factor secretion, or thrombin generation. Thus, endotoxemia induces multi-cellular activation in vivo. Adenosine inhibits leukocyte adhesion and extravasation, and mildly attenuates platelet responsiveness and soluble P-selectin release. Adenosine has the potential of becoming a therapeutic antiinflammatory drug, but an optimal treatment strategy needs to be developed.

 
  • References

  • 1 Marcus AJ. Thrombosis and inflammation as multicellular processes: pathophysiologic significance of transcellular metabolism. Blood 1990; 76: 1903-7.
  • 2 Fredholm BB. Purines and neutrophil leukocytes. General Pharmacol 1997; 28: 345-50.
  • 3 Cronstein BN. Adenosine, an endogenous anti-inflammatory agent. J Appl Physiol 1994; 76: 5-13.
  • 4 Cronstein BN, Kramer SB, Weissmann G, Hirschhorn R. Adenosine: a physiological modulator of superoxide anion generation by human neutrophils. J Exp Med 1983; 158: 1160-77.
  • 5 Bengtsson T, Zalavary S, Stendahl O, Grenegard M. Release of oxygen metabolites from chemoattractant-stimulated neutrophils is inhibited by resting platelets: role of extracellular adenosine and actin polymerization. Blood 1996; 87: 4411-23.
  • 6 Thiel M, Holzer K, Kreimeier U, Moritz S, Peter K, Messmer K. Effects of adenosine on the functions of circulating polymorphonuclear leukocytes during hyperdynamic endotoxemia. Infect Immun 1997; 65: 2136-44.
  • 7 Cronstein BN, Levin RI, Belanoff J, Weissmann G, Hirschhorn R. Adenosine: an endogenous inhibitor of neutrophil-mediated injury to endothelial cells. J Clin Invest 1986; 78: 760-70.
  • 8 Akimitsu T, White JA, Carden DL, Gute DC, Korthuis RJ. Fructose-1,6- diphosphate or adenosine attenuate leukocyte adherence in postischemic skeletal muscle. Am J Physiol 1995; 269: H1734-51.
  • 9 Bouma MG, Stad RK, van den Wildenberg FA, Buurman WA. Differential regulatory effects of adenosine on cytokine release by activated human monocytes. J Immunol 1994; 153: 4159-68.
  • 10 Colli S, Tremoli E. Multiple effects of dipyridamole on neutrophils and mononuclear leukocytes: adenosine-dependent and adenosine-independent mechanisms. J Lab Clin Med 1991; 118: 136-45.
  • 11 Paul S, Feoktistov I, Hollister AS, Robertson D, Biaggioni I. Adenosine inhibits the rise in intracellular calcium and platelet aggregation produced by thrombin: evidence that both effects are coupled to adenylate cyclase. Mol Pharmacol 1990; 37: 870-5.
  • 12 Seligmann C, Kupatt C, Becker BF, Zahler S, Beblo S. Adenosine endogenously released during early reperfusion mitigates postischemic myocardial dysfunction by inhibiting platelet adhesion. J Cardiovasc Pharmacol 1998; 32: 156-63.
  • 13 Edlund A, Siden A, Sollevi A. Evidence for an anti-aggregatory effect of adenosine at physiological concentrations and for its role in the action of dipyridamole. Thromb Res 1987; 45: 183-90.
  • 14 Söderbäck U, Sollevi A, Wallén NH, Larsson PT, Hjemdahl P. Anti-aggregatory effects of physiological concentrations of adenosine in human whole blood as assessed by filtragometry. Clin Sci 1991; 81: 691-4.
  • 15 Nomura F, Aoki M, Forbess JM, Mayer Jr JE. Effects of adenosine infusion with or without leukocyte depletion on recovery after hypothermic ischemia in neonatal lamb hearts. Eur J Cardio-Thorac Surg 1998; 14: 76-81.
  • 16 Sollevi A, Torssell L, Fredholm BB, Settergren G, Blombäck M. Adenosine spares platelets during cardiopulmonary bypass in man without causing systemic vasodilatation. Scand J Thorac Cardiov Surg 1985; 19: 155-9.
  • 17 Mentzer RJ, Rahko PS, Canver CC, Chopra PS, Love RB, Cook TD, Hegge MO, Lasley RD. Adenosine reduces postbypass transfusion requirements in humans after heart surgery. Ann Surg 1996; 224: 523-9.
  • 18 Malhotra R, Priest R, Foster MR, Bird MI. P-selectin binds to bacterial lipopolysaccharide. Eur J Immunol 1998; 28: 983-8.
  • 19 Malhotra R, Bird MI. L-selectin: a novel receptor for lipopolysaccharide and its potential role in bacterial sepsis. Bioessays 1997; 19: 919-23.
  • 20 McCuskey RS, Urbaschek R, Urbaschek B. The microcirculation during endotoxemia. Cardiov Res 1996; 32: 752-63.
  • 21 Holst O, Ulmer AJ, Brade H, Flad HD, Rietschel ET. Biochemistry and cell biology of bacterial endotoxins. FEMS Immunol Med Microbiol 1996; 16: 83-104.
  • 22 Belfrage M, Sollevi A, Segerdahl M, Sjölund K-F, Hansson P. Systemic adenosine infusion alleviates spontaneous and stimulus evoked pain in patients with peripheral neuropathic pain. Anesth Analg 1995; 81: 713-7.
  • 23 Stohlawetz P, Folman CC, von dem Borne AEGK, Pernerstorfer T, Eichler H-G, Panzer S, Jilma B. Effects of endotoxemia on thrombopoiesis in man. Thromb Haemost 1999; 81: 613-7.
  • 24 Pajkrt D, Doran JE, Koster F, Lerch PG, Arnet B, van der Poll T, ten Cate JW, van Deventer SJ. Antiinflammatory effects of reconstituted high-density lipoprotein during human endotoxemia. J Exp Med 1996; 184: 1601-8.
  • 25 Chronos NA, Wilson DJ, Janes SL, Hutton RA, Buller NP, Goodall AH. Aspirin does not affect the flow cytometric detection of fibrinogen binding to, or release of alpha-granules or lysosomes from, human platelets. Clin Sci 1994; 87: 575-80.
  • 26 Li N, Goodall AH, Hjemdahl P. Efficient flow cytometric assay for platelet-leukocyte aggregates in whole blood using fluorescence signal triggering. Cytometry 1999; 35: 154-61.
  • 27 Ault KA, Rinder HM, Mitchell JG, Rinder CS, Lambrew CT, Hillman RS. Correlated measurement of platelet release and aggregation in whole blood. Cytometry 1989; 10: 448-55.
  • 28 Issekutz AC, Ripley M, Jackson JR. Role of neutrophils in the deposition of platelets during acute inflammation. Lab Invest 1983; 49: 716-24.
  • 29 Itoh H, Cicala C, Douglas GJ, Page CP. Platelet accumulation induced by bacterial endotoxin in rats. Thromb Res 1996; 83: 405-19.
  • 30 Michelson AD, Barnard MR, Hechtman HB, MacGregor H, Connolly RJ, Loscalzo J, Valeri CR. In vivo tracking of platelets: circulating degranulated platelets rapidly lose surface P-selectin but continue to circulate and function. Proc Natl Acad Sci USA 1996; 93: 11877-82.
  • 31 Blann AD, Lip GY. Hypothesis: is soluble P-selectin a new marker of platelet activation?. Atheroscler 1997; 128: 135-8.
  • 32 Mannucci PM. von Willebrand factor: a marker of endothelial damage?. Arterioscler Thromb Vascl Biol 1998; 18: 1359-62.
  • 33 Diacovo TG, Puri KD, Warnock RA, Springer TA, von Andrian UH. Platelet-mediated lymphocyte delivery to high endothelial venules. Science 1996; 273: 252-5.
  • 34 Diacovo TG, Roth SJ, Buccola JM, Bainton DF, Springer TA. Neutrophil rolling, arrest, and transmigration across activated, surface-adherent platelets via sequential action of P-selectin and the beta 2-integrin CD11b/CD18. Blood 1996; 88: 146-57.
  • 35 Shi J, Kokubo Y, Wake K. Expression of P-selectin on hepatic endothelia and platelets promoting neutrophil removal by liver macrophages. Blood 1998; 92: 520-8.
  • 36 Li N, Wallén NH, Hjemdahl P. Evidence for prothrombotic effect of exercise and limited protection by aspirin. Circulation 1999; 100: 1374-9.
  • 37 Thiel M, Chambers JD, Chouker A, Fischer S, Zourelidis C, Bardenheuer HJ, Arfors KE, Peter K. Effect of adenosine on the expression of b2 integrins and L-selectin of human polymorphonuclear leukocytes in vitro. J Leuk Biol 1996; 59: 671-82.
  • 38 Dionisotti S, Zocchi C, Varani K, Borea PA, Ongini E. Effects of adenosine derivatives on human and rabbit platelet aggregation. Correlation of adenosine receptor affinities and antiaggregatory activity. N-S Arch Pharmacol 1992; 346: 673-6.
  • 39 Cristalli G, Vittori S, Thompson RD, Padgett WL, Shi D, Daly JW, Olsson RA. Inhibition of platelet aggregation by adenosine receptor agonists. N-S Arch Pharmacol 1994; 349: 644-50.