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Thromb Haemost 2000; 83(02): 327-333
DOI: 10.1055/s-0037-1613807
Rapid Communication
Schattauer GmbH

GPV Is a Marker of In Vivo Platelet Activation – Study in a Rat Thrombosis Model[*]

Catherine Ravanat
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
Monique Freund
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
Pierre Mangin
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
David O. Azorsa
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
Catherine Schwartz
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
Sylvie Moog
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
Simone Schuhler
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
Josiane Dambach
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
Jean-Pierre Cazenave
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
,
François Lanza
From INSERM U.311, Biologie et Pharmacologie de l’Hémostase et de la Thrombose, Etablissement de Transfusion Sanguine de Strasbourg, France
› Author Affiliations
Further Information

Publication History

Received 16 June 1999

Accepted after revision 12 October 1999

Publication Date:
11 December 2017 (online)

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Summary

Thrombin plays a central role in the genesis of thrombotic events and is the most potent known platelet agonist. This enzyme activates platelets by cleaving G-protein coupled protease activated receptors (PARs) and by binding to glycoprotein (GP) Ib. Thrombin also cleaves platelet GPV to liberate a soluble 69 kDa fragment (GPVf1), leaving a 20 kDa fragment (GPVf2) attached to the membrane. The aim of this study was to assess the value of GPV as an in vivo marker of the activation of platelets by thrombin. Newly developed monoclonal and polyclonal antibodies recognizing rat GPVf1 and GPVf2 respectively were used to detect soluble GPV by ELISA and the new NH2-terminus exposed by thrombin using flow cytometry. These assays were employed in a rat thrombosis model designed to trigger thrombin formation in vivo. When thromboplastin (4.8 ml/kg/h) was infused for 30 min, thrombin generation was reflected by a rapid increase in thrombin-antithrombin (TAT) complexes in plasma and by the appearance of GPVf2 at the surface of circulating platelets. Simultaneously, GPVf1 disappeared from the surface of platelets and accumulated as a soluble fragment in plasma, where it was detected by GPV ELISA. These effects were inhibited by pretreatment of the rats with hirudin. Levels of plasma PF4 also increased in this model, but unlike GPV levels which returned slowly (> 2 hours) to baseline, PF4 had a very short half-life.

In conclusion, GPV is cleaved by thrombin in vivo, circulates and is a reliable in vivo marker of the activation of platelets by thrombin. Monitoring of GPV levels in rats should be useful to evaluate the effects of antithrombotic and antiplatelet drugs, while further studies will be required to confirm the potential interest of GPV as a marker of thrombotic states in humans.

Keywords

Rat GPV - thrombin generation - prethrombotic state - animal model - flow cytometry

* Part of this work was presented in preliminary form at the XVIth Congress of the International Society of Thrombosis and Haemostasis, Florence, June 6-12, 1997 and reproduced in abstract form in Thrombosis and Haemostasis, 7: 473, 1997


 

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