The binding of von Willebrand factor (vWf) to stimulated platelets in the plasma milieu was performed using a radiolabeled monoclonal antibody to vWf. Plasma proteins specifically inhibited the thrombin- and ADP/epinephrine-induced vWf binding to activated platelets but did not inhibit the ristocetin-induced vWf binding. When normal plasma was heat defibrinated, monoclonal-labeled vWf was bound to platelets following thrombin or ADP/epinephrine stimulation. Furthermore, monoclonal-labeled vWf from afibrinogenemic plasma bound normally to platelets. The binding of vWf to stimulated platelets in either heat-defibrinated normal plasma or afibrinogenemic plasma was specifically inhibited by the addition of normal plasma fibrinogen in a concentration-dependent manner. At levels of fibrinogen less than 1 mg/ml, however, vWf binding could be demonstrated. The inhibition by fibrinogen of vWf binding to platelets was competitive and overcome by increased concentrations of vWf. These studies show that thrombin-induced and ADP/epinephrine-induced vWf binding to platelets does not occur in the plasma milieu, although at reduced levels of fibrinogen, vWf binding to stimulated platelets can be demonstrated.
J Schullek, J Jordan, R R Montgomery
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