The Use of Monoclonal Antibodies to Human Platelet Membrane Glycoprotein IIb-IIIa to Quantitate Platelet Deposition on Artificial Surfaces

Summary

A new technique is described to quantitate platelet deposition in vitro on artifical surfaces, based on a surface phase radioimmunoassay using the monoclonal antibody 6C9, directed specifically against the membrane glycoprotein complex IIb-IIIa of human platelets. Results correlate in linear fashion with those obtained using ¹¹¹Indium labeled platelets. The method offers particular advantages for the measurement of platelet deposition in whole blood, since platelet separation, washing and labeling procedures are eliminated, together with the ensuing possible selection of platelet populations. In vitro perfusion is performed in glass capillaries of precisely defined diameter (0.80 or 0.56 mm i.d.). Blood flow is laminar and accurately controlled over wall shear rates ranging from venous to capillary (50-4,000 s^-1). Using glass capillaries precoated with purified human albumin or fibrinogen or bovine collagen, platelet deposition from suspensions of washed human platelets in Tyrode's-albumin buffer in the presence of a 40% hematocrit is (platelets/mm²): 11,000 (albumin), 78,000 (fibrinogen) and 306,000 (collagen) after 5 min perfusion at 2,000 s^-1. In heparin, citrate or hirudin anticoagulated whole blood, surfaces are passivated, probably by albumin adsorption from plasma (platelets/mm²): 400 (albumin), 3,600 (fibrinogen) and 48,000 (collagen) after 5 min perfusion in the presence of 13 mM citrate.

Keywords

Platelet deposition - Thrombosis - Monoclonal antibody - Glycoprotein IIb-IIIa - Biomaterials - Capillary perfusion

• References

• 1 Salzman E W, Merrill E W. Interaction of blood with artificial surfaces. In Hemostasis and Thrombosis. Basic Principles and Clinical Practice. Colman R W, Hirsh J, Marder V J, Salzman E W. (eds) J. B. Lippincott Company; Philadelphia: 1982. pp 931-43
• 2 Mustard J F, Packham M A, Kinlough-Rathbone R L. Mechanisms in thrombosis. In Haemostasias and Thrombosis. Bloom A L, Thomas D P. (eds) Churchill Livingstone; Edinburgh: 1981. pp 503-26
• 3 Cazenave J-P. Interaction of platelets with surfaces. In: Blood-Surface Interactions: Biological Principles Underlying Haemocompatibility with Artificial Materials. Cazenave J-P, Davies J A, Kazatchkine M D, van Aken W G. (eds) Elsevier; Amsterdam: 1986. pp 89-105
• 4 Baumgartner H R, Muggli R. Adhesion and aggregation: morphological demonstration and quantitation in vivo and in vitro. In Platelets in Biology and Pathology. Gordon J L. (ed) Elsevier Biomedical Press; Amsterdam: 1976. pp 23-60
• 5 Cazenave J-P, Blondowska D, Richardson M, Kinlough-Rathbone R L, Packham M A, Mustard J F. Quantitative radioisotopic measurement and scanning electron microscopic study of platelet adherence to a collagen-coated surface and to subendothelium with a rotating probe device. J Lab Clin Med 1979; 93: 60-70
• 6 Packham M A, Mustard J F. Platelet adhesion. In Progress in Hemostasis and Thrombosis. Spaet T H. (ed) Grune and Stratton; Orlando, Florida: 1984. Vol 7 pp 211-88
• 7 Cazenave J-P, Hemmendinger S, Beretz A, Sutter-Bay A, Launay J. L’agrégation plaquettaire: outil d’investigation clinique et d’étude pharmacologique. Méthodologie. Ann Biol Clin 1983; 41: 167-79
• 8 Laemmli U K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5
• 9 Kekwick R A, Mackay M E, Nance M H, Record B R. The purification of human fibrinogen. Biochem J 1955; 60: 671-83
• 10 Regoeczi E. The clottability of iodinated fibrinogen. J Nucl Biol Med 1971; 15: 37-41
• 11 Cazenave J-P, Packham M A, Mustard J F. Adherence of platelets to a collagen-coated surface: development of a quantitative method. J Lab Clin Med 1973; 82: 987-90
• 12 Modderman P, Tetteroo P AT, Leeksma O C, Bos M JE, van Mourik J A, Huisman H G, von dem Borne A EG Kr. Differences in functional effects of monoclonal antibodies against platelet membrane glycoproteins Ilb/llla. In Monoclonal Antibodies and Human Blood Platelets: INSERM Symposium N° 27. McGregor J L. (ed) Elsevier; Amsterdam: 1986. pp 131-42
• 13 Regoeczi E. Iodine-Labeled Plasma Proteins. CRC Press; Florida: 1984
• 14 Mustard J F, Perry D W, Ardlie N G, Packham M A. Preparation of suspensions of washed platelets from humans. Brit J Haematol 1972; 22: 193-204
• 15 Eber M, Cazenave J-P, Grob J C, Abecassis J, Methlin G. ¹¹¹Indium labelling of human washed platelets; kinetics and in vivo sequestration sites. In: Blood Cells in Nuclear Medicine, Part I. Cell Kinetics and Bio-distribution. Hardeman M R, Najean Y. (eds) Martinus Nijhoff Publishers; Boston: 1984. pp 29-43
• 16 Goldsmith H L, Turitto V T. Rheological aspects of thrombosis and haemostasis: basic principles and applications. Thromb Haemostas 1986; 55: 415-35
• 17 Sakariassen K S, Baumgartner H S. Axial dependence of platelet deposition on collagen in flowing blood: upstream thrombus growth impairs downstream platelet adhesion. Thromb Haemostas 1985; 54: 109 (Abstr)
• 18 Adams G A, Feuerstein I A. Platelet accumulation on collagen: drugs which inhibit arachidonic acid metabolism and affect intracellular cyclic AMP levels. Thromb Haemostas 1984; 52: 45-9
• 19 Mulvihill J N. Interaction des plaquettes sanguines et des proteines plasmatiques avec des surfaces artificielles. Thesis Doctorat es Sciences Physiques. Université Louis Pasteur de Strasbourg; France: 1984
• 20 Mulvihill J N, Cazenave J-P. Interactions des plaquettes sanguines avec les biomateriaux hemocompatibles. Bull Soc Chim France 1985; 4: 551-4
• 21 Arro E, Collins V P, Brunk U T. High resolution SEM of cultured cells: preparatory procedures. Scanning Electron Microsc 1981; 11: 159-68
• 22 Packham M A, Evans G, Glynn M F, Mustard J F. The effect of plasma proteins on the interaction of platelets with glass surfaces. J Lab Clin Med 1969; 73: 686-97
• 23 Mosher D F. Influence of proteins on platelet-surface interactions. In Interaction of the Blood with Natural and Artificial Surfaces. Salzman E W. (ed) Marcel Dekker; New York: 1981: 85-101
• 24 Zucker M B, Vroman L. Platelet adhesion induced by fibrinogen adsorbed to glass. Proc Soc Exp Biol Med 1969; 131: 318-320
• 25 Brash J L. Hydrophobic polymer surfaces and their interactions with blood. Ann N Y Acad Sci 1977; 283: 356-71
• 26 Turitto V T, Baumgartner H R. Platelet adhesion. In Methods in Hematology: Measurements of Platelet Function. Harker L A, Zimmerman T S. (eds) Vol 8 pp 0046-0063 Churchill Livingstone; Edinburgh: 1983
• 27 Mulvihill J N, Cazenave J-P, Faradji A, Oberling F. Réduction de l’adhésion des plaquettes et de la formation d’un thrombus dans les circuits extracorporels de plasmaphèrése aprés passivation des surfaces par l’albumine. VIIIe Congrés International sur le CEC; Paris: 23-25 juin 1983: 134
• 28 Sigot-Luizard M F, Domurado D, Sigot M, Guidoin R, Gosselin C, Marois M, Girard J F, King M, Badour B. Cytocompatibility of albuminated polyester fabrics. J Biomed Mater Res 1984; 18: 895-909
• 29 Kinlough-Rathbone R L, Mustard J F, Packham M A, Perry D W, Reimers H-J, Cazenave J-P. Properties of washed human platelets. Thromb Haemostas 1977; 37: 291-308
• 30 Baumgartner H R, Muggli R, Tschopp T B, Turitto V T. Platelet adhesion, release and aggregation in flowing blood: Effects of surface properties and platelet function. Thromb Haemostas 1976; 35: 124-38
• 31 Turitto V T, Weiss H J, Baumgartner H R. The effect of shear rate on platelet interaction with subendothelium exposed to citrated human blood. Microvasc Res 1980; 19: 352-65
• 32 Salzman E W, Rosenberg R D, Smith M A, Lindon J N, Faureau L. Effect of heparin and heparin fractions on platelet aggregation. J Clin Invest 1980; 65: 64-73
• 33 Cazenave J-P, Sutter A, Hemmendinger S, Wiesel M-L, Lanza F, Daver J. Adrenaline activates human platelets but does not cause primary aggregation if thrombin generation is inhibited by hirudin. Thromb Haemostas 1981; 46: 95 (Abstr)