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• W2553578997 abstract "Abstract Abstract 1137 Background: Platelets contain in their alpha granules ∼2.5% of the protein S in blood. It has been suggested that this protein S supports the anticoagulant activity of exogenous activated protein C (APC), but it is not known whether protein S that is released from stimulated platelets can exert anticoagulant activity that is independent of APC and TFPI. We recently showed that at least some of the anticoagulant activity of plasma protein S is independent of APC and TFPI, although data suggested that plasma protein S may also have TFPI-dependent activity. Objective and methods: To determine if platelet protein S has anticoagulant activity that is independent of APC and TFPI, prothrombinase and extrinsic FXase reactions were initiated on the surface of fresh stimulated or unstimulated washed platelets in the presence and absence of blocking antibodies against APC, TFPI, and/or protein S, or in the presence and absence of purified plasma-derived protein S. Platelets were adjusted to a concentration of 0.7 to 2 × 10e8/ml, which contained 2.3–6.5 nM total platelet protein S. The last platelet wash contained negligible amounts of plasma protein S. Results: Neutralizing anti-protein S antibodies allowed up to 5.7-fold (mean: 2.1 ± 1.5 –fold, n=13) more thrombin generation on calcium ionophore-stimulated platelets following supplementation with 50–500 pM FXa and 600 nM prothrombin, and allowed up to 2.5-fold (mean: 1.7 ± 0.7 –fold, n=11) more thrombin generation on platelets that were not ionophore-stimulated but were gradually stimulated following FXa and prothrombin supplementation. Anti-protein S antibodies had no effect on thrombin generation on platelets that were treated with prostaglandin E1 (PGE1) to suppress platelet activation and then supplemented with procoagulants. This implies that platelet protein S is released from stimulated platelets and downregulates thrombin generation on platelets, and that neutralizing anti-protein S antibodies block this activity of protein S. Anti-protein S antibodies allowed up to 1.8-fold (mean 1.5 ± 0.2 –fold, n=8) more FXa generation on the surface of stimulated platelets supplemented with 5 pM TF, 100 pM FVIIa, and 160 nM FX, but anti-protein S antibodies had no effect on FXa generation on platelets treated with PGE1. Most of these experiments were performed in the presence of neutralizing antibodies against TFPI and APC, but thrombin and FXa generation on platelets under the varying conditions described were unaffected by the presence of these neutralizing antibodies. Purified plasma-derived zinc-containing protein S downregulated thrombin and FXa generation on platelets (IC50 = 6–18 nM PS) and in plasma >10-fold more potently than zinc-deficient protein S. We could not demonstrate a synergistic anticoagulant effect when TFPI was combined with zinc-deficient protein S in the presence of stimulated platelets and procoagulant proteins. Conclusion: Protein S that is released from stimulated platelets exerts anticoagulant activity that is independent of TFPI and APC. Disclosures: No relevant conflicts of interest to declare." @default.
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• W2553578997 date "2010-11-19" @default.
• W2553578997 modified "2023-10-01" @default.
• W2553578997 title "Protein S Released From Stimulated Platelets Has Anticoagulant Activity Independent of Activated Protein C and Tissue Factor Pathway Inhibitor (TFPI)." @default.
• W2553578997 doi "https://doi.org/10.1182/blood.v116.21.1137.1137" @default.
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