Platelet microparticle membranes have 50- to 100-fold higher specific procoagulant activity than activated platelets

Summary

Platelet microparticles (PMPs) are small vesicles released from blood platelets upon activation. The procoagulant activity of PMPs has been previously mainly characterized by theirability to bind coagulation factors VIII and Va in reconstructed systems. It can be supposed that PMPs can contribute to the development of thrombotic complications in the pathologic states associated with the increase of their blood concentration. In this study we compared procoagulant properties of calcium ionophore A23187-activated platelets and PMPs using several in-vitro models of hemostasis. Surface densities of phosphatidylserine, CD61, CD62P and factor X bound per surface area unit were determined by flow cytometry. They were 2.7-, 8.4-, 4.3-, and 13-fold higher for PMPs than for activated platelets, respectively. Spatial clot growth rate (V_clot) in the reaction-diffus ion experimental model and endogenous thrombin potential (ETP) were determined in plasma, which was depleted of phospholipid cell surfaces by ultra-centrifugation and supplemented with activated platelets or PMPs at different concentrations. Both V_cllot and ETP rapidly increased with the increase of PMP or platelet concentration until saturation was reached. The plateau values of V_clot and ETP for activated platelets and PMPs were similar. In both assays, the procoagulant activity of one PMP was almost equal to that of one activated platelet despite at least two-orders-of-magnitude difference in their surface areas. This suggests that the PMP surface is approximately 50- to 100-fold more procoagulant than the surface of activated platelets.

Keywords

Platelet - PMPs - procoagulant activity - spatial dynamics of coagulation - thrombin generation - factor X - phosphatidylserine - CD-61 - CD-62P

• References

• 1 Schenone M, Furie BC, Furie B. The blood coagulation cascade. Curr Opin Hematol 2004; 11: 272-277.
• 2 Bevers EM, Comfurius P, Dekkers DW. et al. Lipid translocation across the membrane of mammalian cells. Biochim Biophys Acta 1999; 1439: 317-330.
• 3 Kalafatis M, Swords NA, Rand MD. et al. Membrane- dependent reactions in blood coagulation: role of the vitamin K-dependent enzyme complexes. Biochim Biophys Acta 1994; 1227: 113-129.
• 4 Horstman LL, Ahn YS. Platelet microparticles: a wide-angle perspective. Platelet microparticles: a wide-angle perspective. Crit Rev Oncol Hematol 1999; 30: 111-142.
• 5 Freyssinet JM. Cellular microparticles: what are they bad or good for?. J Thromb Haemost 2003; 1: 1655-1662.
• 6 Morel O, Toti F, Hugel B. et al. Cellular microparticles: a disseminated storage pool of bioactive vascular effectors. Curr Opin Hematol 2004; 11: 156-164.
• 7 Ahn YS. Cell-derived microparticles: 'Miniature envoys with many faces'. J Thromb Haemost 2005; 3: 884-887.
• 8 Horstman LL, Jy W, Junenez JJ. et al. New horizons in the analysis of circulating cell-derived microparticles. Keio J Med 2004; 53: 210-230.
• 9 Nieuwland R, Berckmans RJ, Rotteveel-Eijkman RC. et al. Cell-derived microparticles generated in patients during cardiopulmonary bypass is highly procoagulant. Circulation 1997; 96: 3534-3541.
• 10 Nieuwland R, Berckmans RJ, McGregor S. et al. Cellular origin and procoagulant properties of microparticles in meningococcal sepsis. Blood 2000; 95: 930-935.
• 11 Joop K, Berckmans RJ, Nieuwland R. et al. Microparticles from patients with multiple organ dysfunction syndrome and sepsis support coagulation through multiple mechanisms. Thromb Haemost 2001; 85: 810-820.
• 12 Berckmans RJ, Nieuwland R, Böing AN. et al. Cellderived microparticles circulate in healthy humans and support low grade thrombin generation. Thromb Haemost 2001; 85: 639-646.
• 13 Sims PJ, Wiedmer T, Esmon CT. et al. Assembly of platelet prothrombinase complex is linked to vesiculation of platelet plasma membrane. Studies in Scott syndrome: an isolated defect in platelet procoagulant activity. J Biol Chem 1989; 264: 17049-17057.
• 14 Dechary-Prigent J, Freyssinet JM, Pasquet JM. et al. Annexin V as a probe of aminophospholipid exposure and platelet membrane vesiculation: a flow cytometry study showing a role for free sulfhydryl groups. Blood 1993; 81: 2554-2565.
• 15 Sims PJ, Faioni EM, Wiedmer T. et al. Complement proteins C5b-9 cause release of membrane vesicles from the platelet surface that are enriched in the membrane receptor for coagulation factor Va and express prothrombinase activity. J Biol Chem 1988; 263: 18205-18212.
• 16 Gilbert GE, Sims PJ, Wiedmer T. et al. Platelet-derived microparticles express high affinity receptors for factor VIII. J Biol Chem 1991; 266: 17261-17268.
• 17 Tans G, Rosing J, Tomassen MCLGD. et al. Comparison of anticoagulant and procoagulant activities of stimulated platelets and platelet-derived microparticles. Blood 1991; 77: 2641-2648.
• 18 Hemker HC, Giesen PL, Ramjee M. et al. The thrombogram: monitoring thrombin generation in platelet- rich plasma. Thromb Haemost 2000; 83: 589-591.
• 19 Ovanesov MV, Krasotkina JV, Ul'yanova LI. et al. Hemophilia A and B are associated with abnormal spatial dynamics of clot growth. Biochim Biophys Acta 2002; 1572: 45-57.
• 20 Ovanesov MV, Lopatina EG, Saenko EL. et al. Effect of factor VIII on tissue factor-initiated spatial clot growth. Thromb Haemost 2003; 89: 235-242.
• 21 Ovanesov MV, Ananyeva NM, Panteleev MA. et al. Initiation and propagation of coagulation from tissue factor-bearing cell monolayers to plasma: initiator cells do not regulate spatial growth rate. J Thromb Haemost 2005; 3: 321-331.
• 22 Kawabata S, Miura T, Morita T. et al. Highly sensitive peptide-4-methylcoumaryl-7-amide substrates for blood-clotting proteases and trypsin. Eur J Biochem 1988; 172: 17-25.
• 23 Sinauridze EI, Volkova RI, Krasotkina YV. et al. Dynamics of clot growth induced by thrombin diffusing into nonstirred citrate human plasma. Biochim Biophys Acta 1998; 1425: 607-616.
• 24 Korotina NG, Ovanesov MV, Plyushch OP. et al. Study of the spontaneous clots in normal plasma and plasma of haemophilia A patients. Gematol Transfuziol, 2002; 47: 26-29.
• 25 Taube J, McWilliam N, Luddington R. et al. Activated protein C resistance: effect of platelet activation, platelet-derived microparticles, and atherogenic lipoproteins. Blood 1999; 93: 3792-3797.
• 26 Freifelder D. Sedimentation. In: Physical Biochemistry. Applications to Biochemistry and Molecular Biology, Part IV (Hydrodinamic Methods). San Francisco: WH Freeman and Company; 1976: p. 275-349.
• 27 Panteleev MA, Ananyeva NM, Greco NJ. et al. Two subpopulations of thrombin-activated platelets differ in their binding of the components of the intrinsic factor X-activating complex. J Thromb Haemost 2005; 3: 2545-2553.
• 28 Barry OP, Kazanietz MG, Pratico D. et al. Arachidonic acid in platelet microparticles up-regulates cyclooxygenase-2-dependent prostaglandin formation via a protein kinase C/mitogen-activated protein kinase- dependent pathway. J Biol Chem 1999; 274: 7545-7556.
• 29 Vidal C, Spaulding C, Picard F. et al. Flow cytometry detection of platelet procoagulant activity and microparticles in patients with unstable angina treated by percutaneous coronary angioplasty and stent implantation. Thromb Haemost 2001; 86: 784-790.
• 30 Michelson AD, Barnard MR, Krueger LA. et al. Evaluation of platelet function by flow cytometry. Methods 2000; 21: 259-270.
• 31 Sinha D, Seaman FS, Koshy A. et al. Blood coagulation factor XIa binds specifically to a site on activated human platelets distinct from that for factor XI. J Clin Invest 1984; 73: 1550-1556.
• 32 Scandura JM, Ahmad SS, Walsh PN. A binding site expressed on the surface of activated human platelets is shared by factor X and prothrombin. Biochemistry 1996; 35: 8890-8902.
• 33 Muller I, Klocke A, Alex M. et al. Intravascular tissue factor initiates coagulation via circulating microvesicles and platelets. FASEB J 2003; 17: 476-488.
• 34 Hoffman M, Monroe DM, Roberts HR. Coagulation factor IXa binding to activated platelets and platelet- derived microparticles: a flow cytometric study. Thromb Haemost 1992; 68: 74-78.
• 35 Keuren JF, Magdeleyns EJ, Govers-Riemslag JW. et al. Effects of storage-induced platelet microparticles on the initiation and propagation phase of blood coagulation. Br J Haematol 2006; 134: 307-313.
• 36 Fox JE, Austin CD, Reynolds CC. et al. Evidence that agonist-induced activation of calpain causes the shedding of procoagulant-containing microvesicles from the membrane of aggregating platelets. J Biol Chem 1991; 266: 13289-13295.