Measurement of GPV Released by Activated Platelets Using a Sensitive Immunocapture ELISA – Its Use to Follow Platelet Storage in Transfusion

Azorsa, David O.; Moog, Sylvie; Ravanat, Catherine; Schuhler, Simone; Folléa, Gilles; Cazenave, Jean‐Pierre; Lanza, François

doi:10.1055/s-0037-1614430

Cited by 47 publications

(53 citation statements)

References 29 publications

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“…29 Microparticles are the main link between platelet activation and thrombin generation. 29,30 Elevated levels of sGPV and platelet-derived microparticles in vivo indicate platelet activation by thrombin 27 in TAAA. Elevated soluble CD40L (CD154), a transmembrane protein released by proteolytic cleavage in response to various stimuli, 31 in the plasma of TAAA patients also suggests platelet activation.…”

Section: Touat Et Almentioning

confidence: 99%

“…The increased level of TAT is in favor of in vivo thrombin generation, which is further supported by the elevated level of sGPV, a marker of platelet activation by thrombin. 27 The rate of prothrombin activation is dependent on the assembly of tenase and prothrombinase complexes, which in turn depends on the presence of surfaces enriched in negativelycharged phospholipids. 28 The increased number of microparticles of platelet origin observed in association with large aneurysms is likely to provide the phosphatidylserine-enriched membranes required to favor thrombin generation.…”

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confidence: 99%

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Dilation-Dependent Activation of Platelets and Prothrombin in Human Thoracic Ascending Aortic Aneurysm

Touat

Lepage

Ollivier

et al. 2008

ATVB

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Objectives-The purpose of this study was to investigate whether thoracic ascending aortic aneurysm (TAAA) induces platelet activation as mural thrombus participates in aortic dilatation in abdominal aortic aneurysm and TAAA are associated with rheological factors favoring coagulation activation. Methods and Results-We studied the relation between coagulation activation and aortic diameter in Marfan patients (MFS) with various aortic diameters (nϭ52). We then studied patients presenting large aneurysms associated with bicuspid aortic valve (BAV) and degenerative form. Lastly, we used immunochemistry and biochemistry to investigate prothrombin/thrombin retention within the aortic wall. Microparticles, sGPV, tissue factor, and TAT complexes were increased in plasma from MFS with large aneurysms (Ն45 mm) compared to MFS with limited aortic dilatation (Ͻ45 mm). Similar elevations were observed in all patients with large aortic aneurysms, regardless of the etiology, the site of maximal aortic dilation, associated valvulopathy, risk factors, or treatments. P-selectin and platelet-bound fibrinogen were also increased, demonstrating platelet activation in large aneurysms. Significant increase in sCD146 plasma concentration suggested alteration of endothelium. Conclusions-Platelet activation occurs in patients with large aneurysms of the ascending aorta, is dependent on aortic dilation, and is associated with thrombin generation, part of which appears to be retained in mucoid degeneration areas. (Arterioscler Thromb Vasc Biol 2008;28:940-946)Key Words: thoracic aortic aneurysm Ⅲ coagulation Ⅲ Marfan Ⅲ platelets T horacic ascending aortic aneurysms (TAAA) are defined by an enlargement of the ascending aorta. TAAA can be observed in old patients without clear etiologic factors, in whom it represents a primary degenerative pathology of the ascending aorta. 1 But it can also occur in the younger population; it is then usually linked to a genetic or congenital disease. Marfan syndrome represents the majority of these cases. This is usually related to mutations in fibrillin1 2 and rarely to mutations in transforming growth factor (TGF)␤R 2, which can also be responsible for Loeys-Dietz Syndrome and familial aortic aneurysm. [3][4][5] Other molecular abnormalities have been implicated, such as mutations in myosin, 6 TGF␤R 1 , 5 ACTA2, 7 or other currently unknown genes. 8 Moreover, bicuspid aortic valves (BAV), which may be familial, 9 are associated with a high risk of aneurysm of the ascending aorta. 10 In contrast to their etiologic diversity, all localized aortic dilations share common rheological characteristics. In normal human aorta, the regular geometry (tubular shape and parallel walls) results in laminar blood flow and in a homogeneous shear rate, maintaining a physiological steady-state among the blood components and between the circulating blood and arterial wall. 11 Changes in arterial geometry considerably modify rheological parameters and therefore influence interactions between blood components. 11,12 The modu...

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Section: Touat Et Almentioning

confidence: 99%

mentioning

confidence: 99%

Dilation-Dependent Activation of Platelets and Prothrombin in Human Thoracic Ascending Aortic Aneurysm

Touat

Lepage

Ollivier

et al. 2008

ATVB

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“…Similar to other ADAM17 substrates, GPV cleavage occurs near the transmembrane region, leading to the release of an intact GPV ectodomain. The significance of soluble GPV is still unknown, but its generation during platelet activation has been used to develop a specific ELISA for the receptor that may become useful for monitoring platelet activation under conditions of clinical thrombosis (32). This approach was initially based on the hypothesis that thrombin is the major protease that cleaves GPV from the platelet surface, therefore making soluble GPV a specific indicator of intravascular thrombin activity.…”

Section: Fig 4 Gpv Shedding Is Abolished In Adam17mentioning

confidence: 99%

Evidence for a Role of ADAM17 (TACE) in the Regulation of Platelet Glycoprotein V

Rabie

Strehl

Ludwig

et al. 2005

Journal of Biological Chemistry

102

106

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Glycoprotein V (GPV) is a subunit of the GPIb-IX-V receptor for von Willebrand factor and thrombin and has been shown to modulate platelet responses to the two strongest physiological agonists, thrombin and collagen. Thrombin directly cleaves GPV from the platelet surface, yielding a 69-kDa fragment GPV f1 of unknown function. We show here that a ϳ82-kDa fragment of GPV is shed from the platelet surface upon cellular activation with phorbol 12-myristate 13-acetate or the collagen-related peptide. This shedding was inhibited by the broad range metalloproteinase inhibitor GM6001, the two potent ADAM17 inhibitors GW280264X and TAPI-2, and was absent in mice lacking functional ADAM17 (ADAM17 lacking Zn-binding domain; ADAM17 ⌬Zn/⌬Zn ). Furthermore, we show that recombinant ADAM17 ectodomain efficiently releases GPV from the platelet surface. GPV is known to be associated with the intracellular regulatory protein calmodulin, which has previously been shown to be involved in ADAM17-mediated shedding of L-selectin from the surface of leukocytes. As in these reports, inhibition of calmodulin led to rapid GPV shedding from the platelet surface, a process that was again blocked by GM6001 or ADAM17 inhibitors and that was absent in ADAM17 ⌬Zn/⌬Zn mice. Inhibition of outside-in signaling through GPIIb/IIIa did not significantly affect GPV shedding, excluding an essential role of this pathway for the regulation of ADAM17 activity. These results demonstrate that GPV is cleaved upon agonist-induced platelet activation and show that ADAM17 is the major enzyme mediating this process.Platelet adhesion and aggregation at sites of vascular injury is crucial to limit post-traumatic blood loss but may also harm tissue by occluding diseased vessels. The membrane glycoprotein (GP) 1 Ib-IX-V complex plays a central role in these events in that it mediates the initial platelet tethering to the damaged vessel wall under conditions of elevated shear by interacting with collagen-bound von Willebrand factor (1). The receptor complex consists of four distinct polypeptides, GPIb␣ (M r ϭ 143,000), Ib ␤ (M r ϭ 22,000), V (M r ϭ 83,000), and IX (M r ϭ 20,000), in 1:1:0.5:1 stoichiometry with ϳ25,000 copies per platelet. GPs Ib, V, and IX are structurally related and belong to the family of leucine-rich glycoproteins (2). The receptor complex is associated with the regulatory cytoplasmic protein calmodulin, which is involved in the free Ca 2ϩ uptake upon platelet activation (3), but its exact role in GPIb-IX-V function is unclear.The importance of the GPIb-IX-V complex is emphasized by the study of the Bernard-Soulier syndrome, an inherited bleeding disorder in which the complex is congenitally missing or dysfunctional because of mutations in the genes encoding GPIb or GPIX (2). Likewise, targeted deletion of the GPIb␣ (4) or GPIb␤ (5) genes in mice reproduces the Bernard-Soulier phenotype, as characterized by thrombocytopenia, giant platelets, and massively prolonged bleeding times. Although the essential role of GPIb-IX for normal platelet funct...

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“…14,19 The hamster mAb 1C2 against mouse GPV was kindly provided by Dr Junichiro Fujimoto (Tokyo, Japan), 20 and the hamster mAb HM␣2 against mouse ␣2␤1 integrin (CD49b) was from Pharmingen-Becton Dickinson (Le Pont de Claix, France). Human vWF was purified from plasma cryoprecipitates by gel filtration chromatography according to a published procedure, 21 and bovine vWF was kindly provided by Dr S. Jackson, Australian Centre for Blood Diseases (Box Hill, Australia).…”

Section: Reagentsmentioning

confidence: 99%

“…Soluble recombinant GPV was purified from culture media conditioned with GPV-transfected Chinese hamster ovary (CHO) cells using affinity chromatography on a V.1-Sepharose column as previously described. 14 Insoluble bovine collagen type I, soluble human and rat collagen type I, soluble human collagen type III, ADP, the thromboxane analog U46619, PGI 2 , arachidonic acid, and mouse MOPC21 IgG1 were from Sigma Chemicals (St Louis, MO). Equine tendon collagen (a mixture of types I and III) were obtained from Nycomed (Münster, Germany), and human fibrinogen was obtained from Kabi (Stockholm, Sweden).…”

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confidence: 99%

Platelet glycoprotein V binds to collagen and participates in platelet adhesion and aggregation

Moog

Mangin

Lenain

et al. 2001

Blood

Self Cite

125

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Glycoprotein V (GPV) is a subunit of the platelet GPIb-V-IX receptor for von Willebrand factor and thrombin. GPV is cleaved from the platelet surface during activation by thrombin, but its role in hemostasis is still unknown. It is reported that GPV knockout mice had a decreased tendency to form arterial occluding thrombi in an intravital thrombosis model and abnormal platelet interaction with the subendothelium. In vitro, GPV-deficient platelets exhibited defective adhesion to a collagen type I-coated surface under flow or static conditions. Aggregation studies demonstrated a decreased response of the GPV-deficient platelets to collagen, reflected by an increased lag phase and reduced amplitude of aggregation. Responses to adenosine diphosphate, arachidonic acid, and the thromboxane analog U46619 were normal but were enhanced to low thrombin concentrations. The defect of GPV null platelets made them more sensitive to inhibition by the anti-GPVI monoclonal antibody (mAb) JAQ1, and this was also the case in aspirin-or apyrase-treated platelets. Moreover, an mAb (V.3) against the extracellular domain of human GPV selectively inhibited collagen-induced aggregation in human or rat platelets. V.3 injected in rats as a bolus decreased the ex vivo collagen aggregation response without affecting the platelet count. Finally, surface plasmon resonance studies demonstrated binding of recombinant soluble GPV on a collagen-coupled matrix. In conclusion, GPV binds to collagen and appears to be required for normal platelet responses to this agonist. IntroductionPlatelets play a central role in hemostasis through their ability to adhere to a damaged vessel wall and to aggregate in response to agonists such as thrombin, collagen, or adenosine diphosphate (ADP), 1 and these properties are known to be mediated by cell surface glycoproteins. 2,3 However, the exact functions of numerous platelet glycoproteins that have been characterized biochemically remain unknown. Glycoprotein V (GPV, Mr 82 kd) is one of the most abundant glycoproteins at the surface of blood platelets and has long been identified, 4-6 but its functional role is still subject to speculation. GPV is noncovalently linked to the GPIb-IX von Willebrand factor (vWF) receptor on the platelet surface. 7,8 This type I transmembrane protein has a large extracellular domain comprising 15 Leu-rich motifs followed by a thrombin cleavage site. 9,10 The specific release of a soluble 69-kd extracellular domain fragment (GPVf1) by thrombin has led to its proposal as a thrombin receptor, 6 whereas cloning of the gene in rat and mouse revealed a well-conserved thrombin cleavage site. 11 Studies in transfected cells have shown that GPV is required for efficient thrombin binding, possibly through a direct interaction with GPIb␣. 12,13 The release of a soluble fragment by thrombin has been used to develop a specific enzyme-linked immunosorbent assay for GPV that is being tested as a means of monitoring platelet activation under conditions of clinical thrombosis. 14 The role of GPV a...

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Measurement of GPV Released by Activated Platelets Using a Sensitive Immunocapture ELISA – Its Use to Follow Platelet Storage in Transfusion

Cited by 47 publications

References 29 publications

Dilation-Dependent Activation of Platelets and Prothrombin in Human Thoracic Ascending Aortic Aneurysm

Dilation-Dependent Activation of Platelets and Prothrombin in Human Thoracic Ascending Aortic Aneurysm

Evidence for a Role of ADAM17 (TACE) in the Regulation of Platelet Glycoprotein V

Platelet glycoprotein V binds to collagen and participates in platelet adhesion and aggregation

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