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Thromb Haemost 1996; 76(06): 1072-1079
DOI: 10.1055/s-0038-1650708
Original Article
Schattauer GmbH Stuttgart

Detection of Platelet Adhesion/Aggregation to Immobilized Ligands on Microbeads by an Aggregometer

Yoko Minamoto
1   The Department of Internal Medicine, Ehime University School of Medicine, Ehime, Japan
,
Takaaki Hato
2   The Blood Transfusion Division, Ehime University School of Medicine, Ehime, Japan
,
Shingo Nakatani
1   The Department of Internal Medicine, Ehime University School of Medicine, Ehime, Japan
,
Shigeru Fujita
1   The Department of Internal Medicine, Ehime University School of Medicine, Ehime, Japan
› Author Affiliations
Further Information

Publication History

Received 24 April 1996

Accepted after revision 23 August 1996

Publication Date:
11 July 2018 (online)

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Summary

Platelet aggregation is believed to follow platelet adhesion to vascular injury sites. We have developed a turbidimetric assay for platelet aggregation following platelet adhesion to immobilized ligands using an aggregometer. The addition of polystyrene beads coated with von Willebrand factor (vWF) or fibrinogen (Fg) to platelet suspensions caused prompt aggregation of beads and platelets, which was detected as an increase in light transmission. Electron microscopic analysis revealed that platelets adhered to the bead surfaces and that additional platelets adhered to already adhering platelets, leading to the formation of platelet aggregates. vWF-coated beads induced larger aggregates than Fg-coated beads. The interaction of vWF-coated beads with platelets was abolished by both GPIb and GPIIb-IIIa blockers, while that of Fg-coated beads was abolished by GPIIb-IIIa blockers. vWF-coated beads induced modest secretion of granules from platelets but no thromboxane B2 synthesis. Fg-coated beads induced neither reaction. However, pleckstrin phosphorylation and protein tyrosine phosphorylation was induced by both types of bead. Platelet aggregation following platelet adhesion to both types of bead was inhibited by ADP scavengers, a protein kinase C inhibitor and a tyrosine kinase inhibitor, but not by aspirin. These findings suggest that vWF- and Fg-coated beads can induce platelet aggregation following platelet adhesion through specific ligand-receptor interactions and intracellular signaling. Our simple assay using these beads may represent a useful test for immobilized ligand-induced platelet adhesion and aggregation.

 

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