Use of annexin-V to demonstrate the role of phosphatidylserine exposure in the maintenance of haemostatic balance by endothelial cells

Ravanat, Catherine; Archipoff, G; Beretz, Alain; Freund, Gregory G.; Cazenave, J.‐P.; Freyssinet, Jean‐Marie

doi:10.1042/bj2820007

Cited by 67 publications

(56 citation statements)

References 55 publications

(60 reference statements)

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“…4A). The saturating inhibitory concentration of annexin V on phospholipid-dependent processes has previously been shown to be dependent on phospholipid concentration (24). With 0.13 j,g of apoptotic cells per ,ul, half-maximal reversal of the procoagulant effect was observed at 150 ,tM annexin V (Fig.…”

Section: Resultsmentioning

confidence: 90%

Surface blebs on apoptotic cells are sites of enhanced procoagulant activity: implications for coagulation events and antigenic spread in systemic lupus erythematosus.

Casciola‐Rosen

Rosen

Petri

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

450

238

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The restriction of phosphatidylserine (PtdSer) to the inner surface of the plasma membrane bilayer is lost early during apoptosis. Since PtdSer is a potent surface procoagulant, and since there is an increased incidence of coagulation events in patients with systemic lupus erythematosus (SLE) who have anti-phospholipid antibodies, we addressed whether apoptotic cells are procoagulant and whether anti-phospholipid antibodies influence this. Apoptotic HeLa cells, human endothelial cells, and a murine pre-Bcell line were markedly procoagulant in a modified Russell viper venom assay. This procoagulant effect was entirely abolished by addition of the PtdSer-binding protein, annexin V, conrirming that it was PtdSer-dependent. The procoagulant effect was also abolished by addition of IgG purified from the plasma of three patients with anti-phospholipid antibody syndrome, but not IgG from normal controls. Confocal microscopy of apoptotic cells stained with fluorescein-isothiocyanateconjugated-annexin V demonstrated Ca2+-dependent binding to

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Section: Resultsmentioning

confidence: 90%

Surface blebs on apoptotic cells are sites of enhanced procoagulant activity: implications for coagulation events and antigenic spread in systemic lupus erythematosus.

Casciola‐Rosen

Rosen

Petri

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

450

238

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“…1). Endothelial PS exposure supports the assembly of coagulation complexes on the cell surface, leading to thrombin generation 31,32 . Consistent with this, staining of the endothelium with an anti-fibrin monoclonal antibody revealed localized fibrin generation at the sites of needle injury, confirming a-thrombin generation (Fig.…”

Section: Procoagulant Endothelial Cells Recruit Platelets and Leukocymentioning

confidence: 99%

Thrombin-dependent intravascular leukocyte trafficking regulated by fibrin and the platelet receptors GPIb and PAR4

et al. 2015

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Thrombin is a central regulator of leukocyte recruitment and inflammation at sites of vascular injury, a function thought to involve primarily endothelial PAR cleavage. Here we demonstrate the existence of a distinct leukocyte-trafficking mechanism regulated by components of the haemostatic system, including platelet PAR4, GPIba and fibrin. Utilizing a mouse endothelial injury model we show that thrombin cleavage of platelet PAR4 promotes leukocyte recruitment to sites of vascular injury. This process is negatively regulated by GPIba, as seen in mice with abrogated thrombin-platelet GPIba binding (hGPIba D277N ). In addition, we demonstrate that fibrin limits leukocyte trafficking by forming a physical barrier to intravascular leukocyte migration. These studies demonstrate a distinct 'checkpoint' mechanism of leukocyte trafficking involving balanced thrombin interactions with PAR4, GPIba and fibrin. Dysregulation of this checkpoint mechanism is likely to contribute to the development of thromboinflammatory disorders.

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“…This supposition is contradicted by a number of studies, including those showing that endothelial cells, which are not considered procoagulant, nonetheless express significant levels of phosphatidylserine on their surface. 23 This phosphatidylserine is important for maintaining the anticoagulant activity of endothelium by enhancing thrombomodulin activity. 23 A direct comparison of platelets and phospholipids has shown that platelets have complex coagulant activities that are not completely mimicked by phospholipids.…”

Section: Cascade Model Of Coagulationmentioning

confidence: 99%

Platelets and Thrombin Generation

Monroe

Hoffman

Roberts

2002

ATVB

596

479

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Abstract-This review examines the evidence that platelets play a major role in localizing and controlling the burst of thrombin generation leading to fibrin clot formation. From the first functional description of platelets, it has been recognized that platelets supply factors that support the activation of prothrombin. Studies have demonstrated that on activation, the amount of one specific lipid, phosphatidylserine, is significantly increased on the outer leaflet of platelet membranes. When it was found that phosphatidylserine containing lipid extracts could be substituted for platelets in clotting assays, this suggested the possibility that changes in platelet lipid composition were necessary and sufficient to account for platelet surface thrombin generation. Because a growing body of data suggest that platelet-binding proteins provide much of the specificity for platelet thrombin generation, we review in this report data suggesting that changes in lipid composition are necessary but not sufficient to account for platelet surface regulation of thrombin generation. Also, we review data suggesting that platelets from different individuals differ in their capacity to generate thrombin, whereas platelets from a single subject support thrombin generation in a reproducible manner. Individual differences in platelet thrombin generation might be accounted for by differences in platelet-binding proteins.

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Use of annexin-V to demonstrate the role of phosphatidylserine exposure in the maintenance of haemostatic balance by endothelial cells

Cited by 67 publications

References 55 publications

Surface blebs on apoptotic cells are sites of enhanced procoagulant activity: implications for coagulation events and antigenic spread in systemic lupus erythematosus.

Surface blebs on apoptotic cells are sites of enhanced procoagulant activity: implications for coagulation events and antigenic spread in systemic lupus erythematosus.

Thrombin-dependent intravascular leukocyte trafficking regulated by fibrin and the platelet receptors GPIb and PAR4

Platelets and Thrombin Generation

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