title: Platelet microparticle membranes have 50- to 100-fold higher specific procoagulant activity than activated platelets.
abstract: Platelet microparticles (PMPs) are small vesicles released from blood platelets upon activation. The procoagulant activity of PMPs has been previously mainly characterized by their ability 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-diffusion 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(clot) 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.
authors.affiliation: National Research Center for Hematology, Russian Academy of Medical Sciences, 4a Novyi Zykovskii proezd, Moscow 125167, Russia. helen@blood.ru
authors.forename: Elena I
authors.initials: EI
authors.lastname: Sinauridze
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Kireev
authors.forename: Nadezhda Y
authors.initials: NY
authors.lastname: Popenko
authors.forename: Aleksei V
authors.initials: AV
authors.lastname: Pichugin
authors.forename: Mikhail A
authors.initials: MA
authors.lastname: Panteleev
authors.forename: Olga V
authors.initials: OV
authors.lastname: Krymskaya
authors.forename: Fazoil I
authors.initials: FI
authors.lastname: Ataullakhanov
mesh_terms: D001777:Blood Coagulation / Q000187:drug effects; D001780:Blood Coagulation Tests; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D022162:Cytoplasmic Vesicles / Q000378:metabolism*; D005170:Factor X / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D039661:Integrin beta3 / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism*; D007476:Ionophores / Q000494:pharmacology; D016014:Linear Models; D019007:P-Selectin / Q000378:metabolism; D010316:Particle Size; D010718:Phosphatidylserines / Q000378:metabolism; D015539:Platelet Activation / Q000187:drug effects; D013917:Thrombin / Q000378:metabolism; D013997:Time Factors
chemical_list: C061248:ITGB3 protein, human; D039661:Integrin beta3; D007476:Ionophores; D019007:P-Selectin; D010718:Phosphatidylserines; D000001:Calcimycin; D005170:Factor X; D013917:Thrombin; D002118:Calcium