Adhesion and Aggregation of Thrombin Prestimulated Human Platelets on Surface-Bound Fibrinogen: Evidence for Involvement of ADP and Arachidonic Acid Pathways

Summary

The purpose of this study was to examine the influence of substances released from human platelets upon their accumulation on human fibrinogen-coated glass tubes. After prestimulation with thrombin for one minute or in the absence of prestimulation, washed human platelets suspended in Eagle’s medium with RBC were drawn through the tubes at 1 ml/min, 80 s^-1, for 1, 2 or 6 min. Thrombin prestimulation (0.02,0.05 or 0.25 U/ml) was followed by inactivation with hirudin (0.1, 0.25 or 1.25 U/ml) before flow. Singly adherent platelets were observed in the absence of thrombin or with thrombin for exposure times of 1 and 2 min. At 6 min after at least 0.05 U/ml of thrombin, surface- bound aggregates were observed. The initial rate of adhesion increased with the amount of thrombin used for prestimulation. For adhesion to fibrinogen in the absence of prestimulation, platelet-derived ADP was a stimulator. Adhesion was shown to be independent of the ADP and arachidonic acid pathways in response to prestimulation with a low level of thrombin, 0.02 U/ml. For adhesion and cohesion, aggregation, in the presence of sufficient thrombin for prestimulation, 0.05 U/ml, ADP, serotonin and substances from arachidonic acid metabolism acted jointly to stimulate platelets.

Keywords

Platelet adhesion - Platelet aggregation - Fibrinogen-binding - Thrombin

• References

• 1 Adams GA, Feuerstein IA. Kinetics of platelet adhesion and aggregation on protein-coated surfaces: morphology and release from dense granules. asaio Journal 1981; 4: 90-99
• 2 Adams GA, Feuerstein IA. Visual fluorescent and radioisotopic evaluation of platelet accumulation and embolization. Trans Am Soc Artif Intern Organs 1980; 26: 17-22
• 3 Adams GA, Feuerstein IA. Platelet adhesion and release: interfacial concentration of released materials. Am J Physiol 1981; 240: H99-H108
• 4 Adams GA, Feuerstein IA. Maximum fluid concentrations of materials released from platelets at a surface. Am J Physiol 1983; 244: H109-H114
• 5 Feuerstein IA, Dickson VM. Platelet accumulation on fibrinogenblockade of the prostaglandin and ADP pathways. Trans Am Soc Artif Intern Organs 1982; 28: 437-442
• 6 Bevers EM, Comfurius P, Van Rijn JL M L, Hemker HC, Zwaal FA. Generation of prothrombin-converting activity and the exposure of phostatidylserine at the outer surface of platelets. Eur J Biochem 1982; 122: 429-436
• 7 Griffin JH. Surface-dependent activation of blood coagulation. In: Interaction of blood with natural and artificial surfaces. Salzman EW. (Ed) Marcel Dekker; New York: 1982. pp 139-170
• 8 Shuman MA, Majerus PW. The measurement of thrombin in clotting blood by radioimmunoassay. J Clin Invest 1976; 58: 1249-1258
• 9 Horbett TA, Weathersby PK, Hoffman AS. The preferential adsorption of hemoglobin to polyethylene. J Bioengineering 1977; 1: 61-78
• 10 Chuang HY K, King WF, Mason RG. Interaction of plasma proteins with artificial surfaces: protein adsorption isotherms. J Lab Clin Med 1978; 92: 483-496
• 11 Mustard JF, Perry DW, Ardlie NG, Packham MA. Preparation of suspensions of washed platelets from humans. Br J Haematol 1972; 22: 193-204
• 12 Kinlough-Rathbone RL, Mustard JF, Packham MA, Perry DW, Reimers H-J, Cazenave J-P. Properties of washed human platelets. Thromb Haemostas 1977; 37: 291-308
• 13 Lawrie JS, Ross J, Kemp GD. Purification of fibrinogen and separation of its degradation products in the presence of calcium ions. Biochem Soc Trans 1979; 7: 693-694
• 14 Adams GA, Feuerstein IA. How much fibrinogen or fibronectin is enough for platelet adhesion. Trans Am Soc Artif Intern Organs 1981; 27: 219-224
• 15 Cazenave J-P, Packham MA, Mustard JF. Adherence of platelets to a collagen-coated surface: development of a quantitative method. J Lab Clin Med 1973; 82: 978-990
• 16 Sokol R, Rohlf J, Biometry W. Freeman & Co; San Franciso: 1969
• 17 Mustard JF, Packham MA, Kinlough-Rathbone RL, Perry DW, Regoeczi E. Fibrinogen and ADP-induced platelet aggregation. Blood 1978; 52: 453-466
• 18 Di Minno G, Thiagarajan P, Perussia B, Martinez J, Shapiro S, Trinchieri G, Murphy S. Exposure of fibrinogen-binding sites by collagen, arachidonic acid and ADP: inhibition by a monoclonal antibody to the glycoprotein II b-III a complex. Blood 1983; 61: 140-148
• 19 Mustard JF, Kinlough-Rathbone RL, Packham MA, Perry DW, Harfenist EJ, Pai KR M. Comparison of fibrinogen association with normal and thrombasthénie platelets on exposure to ADP or Chymotrypsin. Blood 1979; 54: 987-993
• 20 Niewiarowski S, Budzynski AZ, Morinelli TA, Brudzynski TM, Stewart GJ. Exposure of the fibrinogen receptor on human platelets by proteolytic enzymes. J Biol Chem 1981; 256: 917-925