title: Role of collagen-adherent platelets in mediating fibrin formation in flowing whole blood.
abstract: Activated platelets provide assembly sites for coagulation enzyme complexes and in this way can mediate coagulation during hemostasis and thrombosis. In this study, we examined the procoagulant activity of platelets adhering directly to fibrillar collagen, a main thrombogenic constituent of subendothelium. For this purpose, we used a human ex-vivo thrombosis model in which collagen-coated coverslips were exposed to flowing nonanticoagulated blood (shear rate, 65/s) for 5.5 minutes, which led to the deposition of adherent platelets, platelet thrombi, and fibrin. To examine the procoagulant activity of adherent platelets only, a selective antagonist of the platelet GPIIb-IIIa complex, Ro 44-9883, was infused via a mixing device, resulting in a complete abrogation of platelet thrombus formation but leaving the collagen-adherent platelet layer intact. This platelet layer generated increased postchamber fibrinopeptide A (FPA) levels (203 +/- 33 ng/mL) as compared with control experiments without infusion of inhibitor (95 +/- 13 ng/mL). Concomitantly, fibrin deposition measured by morphometric analysis of cross-sections was also increased, as was the platelet adhesion to collagen. An immunochemical staining of fibrin fibers further showed that the adherent platelets formed the nuclei for fibrin fiber formation. This increase in fibrin deposition was mediated by the intrinsic factor X (F.X) activation complex on adherent single platelets, because almost complete inhibition of FPA generation (9 ng/mL) and fibrin deposition (0.4% +/- 0.2% coverage) was achieved upon coinfusion of the GP IIb-IIIa antagonist and active site-inhibited F.IXa. The large platelet thrombi that were deposited in control experiments contained no significant amounts of immunodetectable fibrin except at the thrombus base, where adherent platelets anchored the thrombi to the collagen surface. These results suggest that the collagen-adherent platelets are important promoters of coagulation during the initial phase of thrombogenesis by providing assembly sites for the F.X activation complex.
authors.affiliation: F. Hoffmann-La Roche Ltd, Pharma Division, Preclinical Research, Basel, Switzerland.
authors.forename: D
authors.initials: D
authors.lastname: Kirchhofer
authors.forename: T B
authors.initials: TB
authors.lastname: Tschopp
authors.forename: B
authors.initials: B
authors.lastname: Steiner
authors.forename: H R
authors.initials: HR
authors.lastname: Baumgartner
mesh_terms: D000085:Acetates / Q000494:pharmacology; D000925:Anticoagulants / Q000494:pharmacology; D001777:Blood Coagulation / Q000187:drug effects / Q000502:physiology; D001792:Blood Platelets / Q000502:physiology*; D003094:Collagen / Q000378:metabolism*; D004789:Enzyme Activation; D015951:Factor Xa / Q000378:metabolism; D005337:Fibrin / Q000096:biosynthesis*; D005344:Fibrinopeptide A / Q000378:metabolism; D018056:Hemorheology; D006801:Humans; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D010477:Perfusion; D010973:Platelet Adhesiveness; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000037:antagonists & inhibitors*; D013927:Thrombosis; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000085:Acetates; D000925:Anticoagulants; D046911:Macromolecular Substances; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D005344:Fibrinopeptide A; D014443:Tyrosine; D005337:Fibrin; D003094:Collagen; C084062:lamifiban; D015951:Factor Xa