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DOI: 10.1055/s-0037-1613789
c7E3 Fab Inhibits Low Shear Flow Modulated Platelet Adhesion to Endothelium and Surface-adsorbed Fibrinogen by Blocking Platelet GP IIb/IIIa as well as Endothelial Vitronectin Receptor
Results from Patients with Acute Myocardial Infarction and Healthy ControlsPublication History
Received
18 May 1999
Accepted after resubmission
15 September 1999
Publication Date:
11 December 2017 (online)
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Summary
The c7E3 Fab reduces ischemic complications in patients undergoing high-risk coronary angioplasty or atherectomy. The present study investigated how c7E3 Fab inhibition of the platelet receptor glycoprotein IIb/IIIa and the endothelial vitronectin receptor affected platelet adhesion to endothelium and surface adsorbed fibrinogen under flow conditions.
Platelet adhesion was examined using a stagnation point flow device with shear stress and shear rates up to 2.2 dynes/cm2 and 170 s−1, respectively. Ex vivo adhesion was compared between two groups of patients with acute myocardial infarction (AMI) treated with angioplasty and stent implantation and a group of healthy controls. Only one AMI group received c7E3 Fab therapy. Patients in both groups were administered acetyl salicylic acid (ASA) and heparin. In AMI patients c7E3 Fab reduced platelet adhesion to adsorbed fibrinogen by 79% compared to AMI patients without c7E3 Fab treatment and by 74% compared to healthy controls. Thirty hours after termination of c7E3 Fab infusion adhesion had slightly recovered with an inhibition of 61% and 52% still present, respectively. Additionally, in vitro platelet adhesion to intact endothelium and to adsorbed fibrinogen was measured during superfusion with ADP stimulated platelet rich plasma of healthy controls to which c7E3 Fab was added at a final concentration fc of 20 µg/ml. In spite of ADP stimulation c7E3 Fab completely blocked platelet adhesion to adsorbed fibrinogen and, moreover, to intact endothelium. Preincubation of endothelial cells with c7E3 (fc = 20 µg/ml) blocked adhesion of ADP-stimulated platelets by approximately 50%.
Apart from the inhibition of platelet aggregation, c7E3 Fab added in vitro and given therapeutically in patients effectively blocks platelet adhesion to components of the injured as well as intact vessel wall under stagnation point flow conditions.
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References
- 1 Mailhac A, Badimon JJ, Fallon JT, Fernandez-Ortiz A, Meyer BJ, Chesebro JH, Fuster V, Badimon L. Effect of an eccentric severe stenosis on fibrin(ogen) deposition on severely damaged vessel wall in arterial thrombosis – relative contribution of fibrin(ogen) and platelets. Circulation 1994; 90: 988-96.
- 2 Wu YP, Van Breugel HHFI, Lankhof H, Wise RJ, Handin RI, De Groot PG, Sixma JJ. Platelet adhesion to multimeric and dimeric von Willebrand factor and to collagen type III preincubated with von Willebrand factor. Arterioscler Thromb Vasc Biol 1996; 16: 611-20.
- 3 Moroi M, Jung SM, Nomura S, Sekiguchi S, Ordinas A, Diaz-Ricart M. Analysis of the involvement of the von Willebrand factor glycoprotein Ib interaction in platelet adhesion to a collagen-coated surface under flow conditions. Blood 1997; 90: 4413-24.
- 4 Reininger AJ, Korndörfer MA, Wurzinger LJ. Adhesion of ADP-activated platelets to intact endothelium under stagnation point flow in vitro is mediated by the integrin αIIbβ3. Thromb Haemost 1998; 79: 998-1003.
- 5 Goldsmith HL, Turitto VT. Rheological aspects of thrombosis and haemostasis: Basic principles and applications. ICTH-report – subcommittee on rheology of the International Committee on Thrombosis and Haemostasis. Thromb Haemost 1986; 55: 415-35.
- 6 Petschek H, Adamis D, Kantrowitz AR. Stagnation flow thrombus formation. Trans Amer Soc Artif Intern Organs 1968; XIV: 256-60.
- 7 Reininger CB, Graf J, Reininger AJ, Spannagl M, Steckmeier B, Schweiberer L. Increased platelet and coagulatory activity indicate ongoing thrombogenesis in peripheral arterial disease. Thromb Res 1996; 82: 523-32.
- 8 Topol EJ. Prevention of cardiovascular ischemic complications with new platelet glycoprotein IIb/IIIa inhibitors. Am Heart J 1995; 130 Suppl. 666-72.
- 9 The EPILOG Investigators. Platelet glycoprotein IIb/IIIa receptor blockade and low-dose heparin during percutaneous coronary revascularization. New Engl J Med 1997; 336: 1689-96.
- 10 Ruggeri ZM. Mechanisms of shear-induced platelet adhesion and aggregation. Thromb Haemost 1993; 70: 119-23.
- 11 Savage B, Saldívar E, Ruggeri ZM. Initiation of platelet adhesion by arrest onto fibrinogen or translocation on von Willebrand factor. Cell 1996; 84: 289-97.
- 12 Reininger AJ, Reininger CB, Spannagl M, Mellinghoff A, Porr A, Heinzmann U, Wurzinger LJ. Effect of fibrinogen substitution in afibrinogenemia on hemorheology and platelet function. Thromb Haemost 1995; 74: 853-8.
- 13 Reininger CB, Reininger AJ, Graf J, Eibl-Eibesfeldt BI, Steckmeier B. Real-time analysis of platelet adhesion under stagnation point flow conditions: The effect of red blood cells and glycoprotein IIb/IIIa receptor blockade. J Vasc Invest 1996; 02: 1-11.
- 14 Cheresh DA, Berliner SA, Vicente V, Ruggeri ZM. Recognition of distinct adhesive sites on fibrinogen by related integrins on platelet and endothelial cells. Cell 1989; 58: 945-53.
- 15 Jordan RE, Wagner CL, Mascelli MA, Treacy G, Nedelman MA, Woody JN, Weisman HF, Coller BS. Preclinical development of c7E3 Fab; a mouse/human chimeric monoclonal antibody fragment that inhibits platelet function by blockade of GPIIb/IIIa receptors with observations on the immunogenicity of c7E3 Fab in humans. In: Horton MA. ed. Adhesion Receptors as Therapeutic Targets. Boca Raton, New York, London, Tokyo: CRC Press; 1996: 281-305.
- 16 Turner NA, Moake JL, Kamat SG, Schafer AI, Kleiman NS, Jordan R, McIntire LV. Comparative real-time effects on platelet adhesion and aggregation under flowing conditions of in vivo aspirin, heparin, and monoclonal antibody fragment against glycoprotein IIb-IIIa. Circulation 1995; 91: 1354-62.
- 17 Coller BS, Anderson K, Weisman HF. New antiplatelet agents: Platelet GPIIb/IIIa antagonists. Thromb Haemost 1995; 74: 302-8.
- 18 Reverter JC, Béguin S, Kessels H, Kumar R, Hemker HC, Coller BS. Inhibition of platelet-mediated, tissue factor-induced thrombin generation by the mouse/human chimeric 7E3 antibody – Potential implications for the effect of c7E3 Fab treatment on acute thrombosis and “clinical restenosis”. J Clin Invest 1996; 98: 863-74.
- 19 Reininger AJ, Heinzmann U, Reininger CB, Friedrich P, Wurzinger LJ. Flow mediated fibrin thrombus formation in an endothelium-lined model of an arterial branching. Thromb Res 1994; 74: 629-41.
- 20 Thiruvikraman SV, Guha A, Roboz J, Taubman MB, Nemerson Y, Fallon JT. In situ localization of tissue factor in human atherosclerotic plaques by binding of digoxigenin-labeled factors VIIa and X. Lab Invest 1996; 75: 451-61.
- 21 Hantgan RR, Hindriks G, Taylor RG, Sixma JJ, De Groot PG. Glycoprotein Ib, von Willebrand factor, and glycoprotein IIb/IIIa are all involved in platelet adhesion to fibrin in flowing whole blood. Blood 1990; 76: 345-53.
- 22 Zaidi TN, McIntire LV, Farrell DH, Thiagarajan P. Adhesion of platelets to surface-bound fibrinogen under flow. Blood 1996; 88: 2967-72.
- 23 Perkkio J, Wurzinger LJ, Schmid-Schonbein H. Fahraeus-Vejlens effect: margination of platelets and leukocytes in blood flow through branches. Thromb Res 1988; 50: 357-64.
- 24 Horbett TA. Principles underlying the role of adsorbed plasma proteins in blood interactions with foreign materials. Cardiovasc Pathol 1993; 02: 137S-48S.
- 25 Wojciechowski P, Brash JL. The Vroman effect in tube geometry: the influence of flow on protein adsorption measurements. J Biomater Sci Polymer Edn 1991; 02: 203-16.
- 26 Stanford MF, Munoz PC, Vroman L. Platelets Adhere where Flow has Left Fibrinogen on Glass. Ann NY Acad Sci 1983; 416: 504-12.
- 27 Hantgan RR, Endenburg SC, Cavero I, Marguerie G, Uzan A, Sixma JJ, De Groot PG. Inhibition of platelet adhesion to fibrin(ogen) in flowing whole blood by Arg-Gly-Asp and fibrinogen gamma-chain carboxy terminal peptides. Thromb Haemost 1992; 68: 694-700.
- 28 Konstantopoulos K, Kamat SG, Schafer AI, Banez EI, Jordan R, Kleiman NS, Hellums JD. Shear-induced platelet aggregation is inhibited by in vivo infusion of an anti-glycoprotein IIb/IIIa antibody fragment, c7E3 Fab, in patients undergoing coronary angioplasty. Circulation 1995; 91: 1427-31.
- 29 Kleiman NS, Raizner AE, Jordan R, Wang AL, Norton D, Mace KF, Joshi A, Coller BS, Weisman HF. Differential inhibition of platelet aggregation induced by adenosine diphosphate or a thrombin receptor-activating peptide in patients treated with bolus chimeric 7E3 Fab: Implications for inhibition of the internal pool of GPIIb/IIIa receptors. J Am Coll Cardiol 1995; 26: 1665-71.
- 30 Coller BS. Blockade of platelet GPIIb/IIIa receptors as an antithrombotic strategy. Circulation 1995; 92: 2373-80.
- 31 Coller BS, Lang D, Scudder LE. Rapid and simple platelet function assay to assess glycoprotein IIb/IIIa receptor blockade. Circulation 1997; 95: 860-7.
- 32 Reininger AJ, Reininger CB, Wurzinger LJ. The influence of fluid dynamics upon adhesion of ADP-stimulated human platelets to endothelial cells. Thromb Res 1993; 71: 245-9.
- 33 Bombeli T, Schwartz BR, Harlan JM. Adhesion of activated platelets to endothelial cells: evidence for a GPIIb/IIIa-dependent bridging mechanism and novel roles for endothelial intercellular adhesion molecule 1 (ICAM-1), αvβ3 integrin, and GPIbα. J Exp Med 1998; 187: 329-39.
- 34 Gawaz M, Neumann FJ, Dickfeld T, Reininger AJ, Adelsberger H, Gebhardt A, Schömig A. Vitronectin receptor (αv β3) mediates platelet adhesion to the luminal aspect of endothelial cells. Implications for reperfusion in acute myocardial infarction. Circulation 1997; 96: 1809-18.