Adhesion of platelets to purified solid-phase von Willebrand factor: effects of wall shear rate, ADP, thrombin, and ristocetin.
When platelets are stimulated with adenosine diphosphate (ADP), thrombin, or ristocetin, they bind soluble von Willebrand factor (vWF). In contrast, platelets adhere to solid-phase vWF without apparent stimulus. This work characterizes the adhesion of washed human platelets to highly purified solid-phase human vWF. VWF (iodine 125-labeled vWF) was demonstrated to bind in a quantifiable fashion to the internal surfaces of glass capillary tubes, saturating at a surface density of 3.0 mg/ml. The multimeric structure of bound vWF was the same as that of normal vWF. Platelets were washed, labeled with indium 111, and resuspended with washed red blood cells (RBCs) in balanced salt solution containing Ca++, Mg++, and apyrase. The washed platelet RBC suspension was aspirated through capillary tubes to which vWF was adsorbed. Adhesion of platelets to adsorbed vWF was directly dependent on the surface density of vWF. Increasing wall shear rate (100 to 5000 sec-1) produced increasing platelet adhesion to maximum reached at 2500 sec-1. Platelets bound to the solid-phase vWF in an irreversible fashion, and, as demonstrated with scanning electron microscopy, they spread on the surface. When used to stimulate the platelets, ADP, thrombin, and ristocetin all increased the platelet adhesion to solid-phase vWF. ADP- and thrombin-stimulated reactions were inhibited by prior treatment of the platelets with 5'-p-fluorosulfonylbenzoyl adenosine. This inhibitor of ADP binding had no effect on the baseline platelet adhesion reaction (without ADP or thrombin). Adenosine in concentration up to 1 mmol/L failed to inhibit adhesion. The data demonstrate that washed platelets adhere to solid-phase vWF without added agonists, that the reaction is dependent on surface density vWF and wall shear rate, that they bind irreversibly, and that they demonstrate surface spreading. In addition, these platelets can be stimulated to increase their adherence to vWF by using ADP, thrombin, and ristocetin.
Olson
J D
JD
Department of Pathology, University of Iowa College of Medicine, Iowa City 52242.
Zaleski
A
A
Herrmann
D
D
Flood
P A
PA
D000244:Adenosine Diphosphate / Q000494:pharmacology*; D000327:Adsorption; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D005898:Glass; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D008855:Microscopy, Electron, Scanning; D008970:Molecular Weight; D010973:Platelet Adhesiveness / Q000187:drug effects; D012016:Reference Values; D012310:Ristocetin / Q000494:pharmacology*; D013917:Thrombin / Q000502:physiology*; D014841:von Willebrand Factor / Q000302:isolation & purification / Q000502:physiology*
D046911:Macromolecular Substances; D014841:von Willebrand Factor; D012310:Ristocetin; D000244:Adenosine Diphosphate; D013917:Thrombin