Reactivity of human platelets with immobilized fibrinogen is dictated by the chemical character of the surface.
Quiescent platelets readily adhere to surface-immobilized fibrinogen. In contrast, platelets exposed to soluble fibrinogen do not demonstrate such activity. As part of an effort to characterize this phenomenon, a solid-phase reagent was prepared by adsorbing human fibrinogen to polystyrene-divinylbenzene microparticles. Using a suspension of human platelets, phase-contrast microscopy was used to quantitate directly platelets bound to fibrinogen-coated beads. This method is fast, straightforward, and requires minimal amounts of reagents and sample. An existing turbidimetric assay was modified to monitor optically the rate and extent of platelet-fibrinogen binding. When platelet-rich plasma was added to a stirred suspension of fibrinogen-coated beads, the rate of aggregation was related directly to the concentration of fibrinogen on the bead surface. This response could not be mitigated by the thrombin inhibitor, hirudin, indicating that any thrombin generated in the reaction has no role in bead aggregation. Conversely, the alpha(IIb)beta(3) antagonist, abciximab (ReoPro), completely prevented aggregation, implicating specific fibrinogen-alpha(IIb)beta(3) interactions as responsible for the observed effect. Beads coated with either albumin or a densely packed, pure film of the neutral phospholipid, phosphatidylcholine (lecithin), do not aggregate under identical conditions, nor do fibrinogen-coated beads aggregate when platelet-depleted plasma is added. When fibrinogen was coated to beads as a mixed film with lecithin, a striking increase in reactivity toward platelets was demonstrated, compared to unmodified beads. These studies indicate that the observed adhesion of platelets to beads is a direct result of platelet-fibrinogen interactions and platelets respond differently to fibrinogen when presented as a mixed film with lipid, compared to the protein alone at an interface.
Cook
B C
BC
Clinical Laboratory Science, Marquette University, PO Box 1881, Milwaukee, WI 53201, USA. bccook@beckman.com
D004305:Dose-Response Relationship, Drug; D005340:Fibrinogen / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008567:Membranes, Artificial; D008863:Microspheres; D009391:Nephelometry and Turbidimetry; D010713:Phosphatidylcholines / Q000494:pharmacology; D010973:Platelet Adhesiveness / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000494:pharmacology; D013499:Surface Properties; D013917:Thrombin / Q000494:pharmacology
D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D005340:Fibrinogen; D013917:Thrombin