Cell-derived Microparticles Circulate in Healthy Humans and Support Low Grade Thrombin Generation

We determined the numbers, cellular origin and thrombin-generating properties of microparticles in healthy individuals (n = 15). Microparticles, isolated from fresh blood samples and identified by flow cytometry, originated from platelets [237 x 10(6)/L (median; range 116-565)], erythrocytes (28 x 10(6)/L; 13-46), granulocytes (46 x 10(6)/L; 16-94) and endothelial cells (64 x 10(6)/L; 16-136). They bound annexin V, indicating surface exposure of phosphatidylserine, and supported coagulation in vitro. Interestingly, coagulation occurred via tissue factor (TF)-independent pathways, because antibodies against TF or factor (F)VII were ineffective. In contrast, in our in vitro experiments coagulation was partially inhibited by antibodies against FXII (12%, p = 0.006), FXI (36%, p <0.001), FIX (28%, p <0.001) or FVIII (32%, p <0.001). Both the number of annexin V-positive microparticles present in plasma and the thrombin-generating capacity inversely correlated to the plasma concentrations of thrombin-antithrombin complex (r = -0.49, p = 0.072 and r = -0.77, p = 0.001, respectively), but did not correlate to prothrombin fragment F1+2 (r = -0.002, p = 0.99). The inverse correlations between the number of microparticles and their thrombin-forming capacity and the levels of thrombin-antithrombin complex in plasma may indicate that microparticles present in the circulation of healthy individuals have an anticoagulant function by promoting the generation of low amounts of thrombin that activate protein C. We conclude that microparticles in blood from healthy individuals support thrombin generation via TF- and FVII-independent pathways, and which may have an anticoagulant function.

[1]  N. Mackman,et al.  Measurement of Tissue Factor Activity in Whole Blood , 2000, Thrombosis and Haemostasis.

[2]  F. Rosendaal,et al.  High levels of coagulation factor XI as a risk factor for venous thrombosis. , 2000, The New England journal of medicine.

[3]  R. Westendorp,et al.  Cellular origin and procoagulant properties of microparticles in meningococcal sepsis. , 2000, Blood.

[4]  G. Grau,et al.  In vitro generation of endothelial microparticles and possible prothrombotic activity in patients with lupus anticoagulant. , 1999, The Journal of clinical investigation.

[5]  J. Badimón,et al.  Blood-borne tissue factor: another view of thrombosis. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[6]  S. Nakamura,et al.  Expression of functional tissue factor on small vesicles of lipopolysaccharide-stimulated human vascular endothelial cells. , 1998, Thrombosis research.

[7]  M. Nishikawa,et al.  Effect of Cilostazol on Soluble Adhesion Molecules and Platelet-derived Microparticles in Patients with Diabetes , 1998, Thrombosis and Haemostasis.

[8]  O. Bagasra,et al.  Molecular Cloning of Platelet Factor XI, an Alternative Splicing Product of the Plasma Factor XI Gene* , 1998, The Journal of Biological Chemistry.

[9]  P. Walsh,et al.  Tissue-specific expression of functional platelet factor XI is independent of plasma factor XI expression. , 1998, Blood.

[10]  P. Arck,et al.  Cutting Edge: Cytokine-Dependent Abortion in CBA × DBA/2 Mice Is Mediated by the Procoagulant fgl2 Prothombinase , 1998, The Journal of Immunology.

[11]  P. Arck,et al.  Cytokine-dependent abortion in CBA x DBA/2 mice is mediated by the procoagulant fgl2 prothrombinase [correction of prothombinase]. , 1998, Journal of immunology.

[12]  B. Osterud.  Tissue factor expression by monocytes: regulation and pathophysiological roles. , 1998, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[13]  K. Maquelin,et al.  Cell-derived microparticles generated in patients during cardiopulmonary bypass are highly procoagulant. , 1997, Circulation.

[14]  L. Horstman,et al.  Platelet microparticles and calcium homeostasis in acute coronary ischemias , 1997, American journal of hematology.

[15]  P. Walsh,et al.  Annexin V inhibition of factor IXa-catalyzed factor X activation on human platelets and on negatively-charged phospholipid vesicles. , 1996, Biochemistry.

[16]  T. Warkentin.  Heparin-induced thrombocytopenia: IgG-mediated platelet activation, platelet microparticle generation, and altered procoagulant/anticoagulant balance in the pathogenesis of thrombosis and venous limb gangrene complicating heparin-induced thrombocytopenia. , 1996, Transfusion medicine reviews.

[17]  P. Siljander,et al.  Platelet-derived microparticles associate with fibrin during thrombosis. , 1996, Blood.

[18]  J. Kelton,et al.  Isolation and characterization of cysteine proteinase in thrombotic thrombocytopenic purpura , 1996, British journal of haematology.

[19]  F. Taylor,et al.  Role of tissue factor and factor VIIa in the coagulant and inflammatory response to LD100 Escherichia coli in the baboon. , 1996, Haemostasis.

[20]  P. A. von dem Borne,et al.  Feedback activation of factor XI by thrombin in plasma results in additional formation of thrombin that protects fibrin clots from fibrinolysis. , 1995, Blood.

[21]  P. Holme,et al.  Platelet‐derived microvesicles and activated platelets express factor Xa activity , 1995, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[22]  H. Shiku,et al.  Tissue factor expression in endothelial cells in health and disease , 1995, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[23]  J. Kelton,et al.  Sera from patients with heparin-induced thrombocytopenia generate platelet-derived microparticles with procoagulant activity: an explanation for the thrombotic complications of heparin-induced thrombocytopenia. , 1994, Blood.

[24]  H. Hemker,et al.  A Chromogenic Test to Determine the Procoagulant Phospholipids in Platelet-rich Plasma and Whole Blood , 1994, Thrombosis and Haemostasis.

[25]  Y. Komiyama,et al.  Amyloid β-Protein Precursor-Rich Platelet Microparticles in Thrombotic Disease , 1994, Thrombosis and Haemostasis.

[26]  J. Freyssinet,et al.  Monocyte vesiculation is a possible mechanism for dissemination of membrane-associated procoagulant activities and adhesion molecules after stimulation by lipopolysaccharide. , 1994, Journal of immunology.

[27]  P. A. von dem Borne,et al.  Surface Independent Factor XI Activation by Thrombin in the Presence of High Molecular Weight Kininogen , 1994, Thrombosis and Haemostasis.

[28]  A. Shibata,et al.  Tissue factor in plasma of patients with disseminated intravascular coagulation , 1994, American journal of hematology.

[29]  P. Holland,et al.  Prolonged circulation of activated platelets following plasmapheresis , 1994, Journal of clinical apheresis.

[30]  L. Horstman,et al.  Elevated platelet microparticles in transient ischemic attacks, lacunar infarcts, and multiinfarct dementias. , 1993, Thrombosis research.

[31]  C. Esmon,et al.  Antithrombotic effects of thrombin-induced activation of endogenous protein C in primates. , 1993, The Journal of clinical investigation.

[32]  M. Levi,et al.  The activation of factor X and prothrombin by recombinant factor VIIa in vivo is mediated by tissue factor. , 1993, The Journal of clinical investigation.

[33]  R. Colman,et al.  The contact system contributes to hypotension but not disseminated intravascular coagulation in lethal bacteremia. In vivo use of a monoclonal anti-factor XII antibody to block contact activation in baboons. , 1993, The Journal of clinical investigation.

[34]  M. Nishikawa,et al.  Significance of platelet-derived microparticles in uremia. , 1993, Nephron.

[35]  B. Lämmle,et al.  Factor XII clotting activity and antigen levels in patients with thromboembolic disease , 1992, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[36]  C. Hack,et al.  A Novel Sensitive Assay for Functional Factor XII Based on the Generation of Kallikrein-C1-lnhibitor Complexes in Factor Xll-Deficient Plasma by Glass-Bound Factor XII , 1992, Thrombosis and Haemostasis.

[37]  P. Mannucci,et al.  Factor IXa-factor VIIIa-cell surface complex does not contribute to the basal activation of the coagulation mechanism in vivo. , 1992, Blood.

[38]  L. Horstman,et al.  Clinical significance of platelet microparticles in autoimmune thrombocytopenias. , 1992, The Journal of laboratory and clinical medicine.

[39]  P. Mannucci,et al.  No activation of the common pathway of the coagulation cascade after a highly purified factor IX concentrate , 1991, British journal of haematology.

[40]  B. Furie,et al.  Platelet-derived microparticles express high affinity receptors for factor VIII. , 1991, The Journal of biological chemistry.

[41]  G. Broze,et al.  Factor XI activation in a revised model of blood coagulation , 1991, Science.

[42]  J. Griffin,et al.  Comparison of anticoagulant and procoagulant activities of stimulated platelets and platelet-derived microparticles. , 1991, Blood.

[43]  B. Lämmle,et al.  Thromboembolism and Bleeding Tendency in Congenital Factor XII DeficienCy - A Study on 74 Subjects from 14 Swiss Families , 1991, Thrombosis and Haemostasis.

[44]  F. Taylor,et al.  Lethal E. coli septic shock is prevented by blocking tissue factor with monoclonal antibody. , 1991, Circulatory shock.

[45]  H. ten Cate,et al.  Factor IX is activated in vivo by the tissue factor mechanism. , 1990, Blood.

[46]  M. Bednarek,et al.  DETECTION OF FACTOR X ACTIVATION IN HUMANS , 1989, Thrombosis and Haemostasis.

[47]  P. Walsh,et al.  Platelet receptor-mediated factor X activation by factor IXa. High-affinity factor IXa receptors induced by factor VIII are deficient on platelets in Scott syndrome. , 1989, The Journal of clinical investigation.

[48]  H. Hemker,et al.  The Effect of Trace Amounts of Tissue Factor on Thrombin Generation in Platelet Rich Plasma, its Inhibition by Heparin , 1989, Thrombosis and Haemostasis.

[49]  P. Sims,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. , 1988, The Journal of biological chemistry.

[50]  D. Altieri,et al.  The saturable high affinity association of factor X to ADP-stimulated monocytes defines a novel function of the Mac-1 receptor. , 1988, The Journal of biological chemistry.

[51]  H. Hauser,et al.  Single bilayer vesicles prepared without sonication. Physico-chemical properties. , 1976, Biochimica et biophysica acta.