Subscribe to RSS
DOI: 10.1160/TH03-01-0027
Thrombin generation in severe haemophilia A and B: the endogenous thrombin potential in platelet-rich plasma
Publication History
Received
16 January 2003
Accepted after revision
02 June 2003
Publication Date:
05 December 2017 (online)
Summary
Thrombin generation was investigated in platelet-rich plasma (PRP) from 11 healthy controls, 17 patients with severe haemophilia A and 7 patients with severe haemophilia B. Mean endogenous thrombin potential (ETP) in arbitrary fluorescence units (FU) was 226.9 ± 44.6, 186.4 ± 22.5, 154.2 ± 41.3 in controls, haemophilia A and B, respectively, all at a platelet count of 200 × 109/l (p = 0.004 for controls vs.haemophilia A, p = 0.003 for controls vs. haemophilia B, no significant difference between haemophilia A and B). The contribution of FVIII to thrombin generation in haemophilia A was 1.31 ± 0.16 FU/% of FVIII:C activity , while for FIX in haemophilia B this was 0.80 ± 0.21 FU/% of FIX activity. There was an almost linear relationship between increasing platelet count and thrombin generation up to a mean platelet count of 100 × 109/l. Further increase in platelet count has only a marginal influence on thrombin generation. Platelets increase ETP in haemophilia A by 0.184 ± 0.022 FU/109platelets/l and in haemophilia B by 0.319 ± 0.085 FU/109platelets/l, and this was significantly different between the two groups (p = 0.0002). This influence of platelets diminishes with increasing concentration of either FVIII or FIX. In conclusion, there is a difference in thrombin generation between haemophilia A and B, and this may be attributed to the role of platelets in the assembly of the tenase complex on their surface.
-
References
- 1 Davie EW, Ratnoff OD. Waterfall sequence for intrinsic blood clotting. Science 1964; 145: 1310-12.
- 2 Hoffman M, Monroe DM. A cell-based model of hemostasis. Thromb Haemost 2001; 85: 958-65.
- 3 Heemskerk JWM, Bevers EM, Lindhout T. Platelet activation and blood coagulation. Thromb Haemost 2002; 88: 186-93.
- 4 Rosing J, van Rijn JL, Bevers EM. et al The role of activated human platelets in prothrom-bin and factor X activation. Blood 1985; 65: 319-32.
- 5 Tans G, Rosing J, Thomassen MC. et al Comparison of anticoagulant and procoagulant activities of stimulated platelets and platelet-derived microparticles. Blood 1991; 77: 2641-8.
- 6 Berckmans RJ, Nieuwland R, BÖing AN. et al Cell-derived microparticles circulate in healthy humans and support low grade throm-bin generation. Thromb Haemost 2001; 85: 639-46.
- 7 Moyer MP, Tracy RP, Tracy PB. et al Plasma lipoproteins support prothrombinase and other procaogulant enzymatic complexes. Arterioscler Thromb Vasc Biol 1998; 18: 458-65.
- 8 Stern D, Nawroth P, Handley D. et al An endothelial cell-dependent pathway of coagulation. Proc Natl Acad Sci USA 1985; 82: 2523-7.
- 9 Peyrou V, Lormeau JC, Hérault JP. et al Contribution of erythrocytes to thrombin generation in whole blood. Thromb Haemost 1999; 81: 400-6.
- 10 Tracy PB, Eide LL, Mann KG. Human pro-thrombinase complex assembly and function on isolated peripheral blood cell populations. J Biol Chem 1985; 260: 2119-24.
- 11 Gailani D, Broze Jr GJ. Factor XI activation in a revised model of blood coagulation. Science 1991; 253: 909-12.
- 12 Pieters J, Lindhout T, Hemker HC. In situ-generated thrombin is the only enzyme that effectively activates factor VIII and factor V in thromboplastin-activated plasma. Blood 1989; 74: 1021-4.
- 13 Butenas S, vant Veer C, Mann KG. Evaluation of the initiation phase of blood coagulation using ultrasensitive assays for serine proteases. J Biol Chem 1997; 272: 21527-33.
- 14 Hemker HC, Wielders S, Kessels H. et al Continuous registration of thrombin generation in plasma, its use for the determination of the thrombin potential. Thromb Haemost 1993; 71: 617-24.
- 15 Bendetowicz AV, Kai H, Knebel R. et al The effect of subcutaneous injection of unfractionated and low molecular weight heparin on thrombin generation in platelet rich plasma – a study in human volunteers. Thromb Haemost 1994; 72: 705-12.
- 16 Hemker HC, Béguin S. Phenotyping the clotting system. Thromb Haemost 2000; 84: 747-51.
- 17 Kakkar VV, Hoppenstead DA, Fareed J. et al Randomized trial of different regimens of heparins in vivo thrombin generation in acute deep vein thrombosis. Blood 2002; 99: 1965-70.
- 18 Keularts IM, Béguin S, de Zwaan C. et al Treatment with GPIIb/IIIa antagonist inhibits thrombin generation in platelet rich plasma from patients. Thromb Haemost 1998; 80: 370-1.
- 19 Hérault JP, Peyrou V, Savi P. et al Effect of SR121566A, a potent GPIIb-IIIa antagonist on platelet-mediated thrombin generation in vitro and in vivo. Thromb Haemost 1998; 79: 383-8.
- 20 Hérault JP, Dol F, Gaich C. et al Effect of clopidogrel on thrombin generation in platelet-rich plasma in the rat. Thromb Haemost 1999; 81: 957-60.
- 21 Wegert W, Graff J, Kaiser D. et al Effects of antiplatelet agents on platelet-induced throm-bin generation. Int J Clin Pharmacol Ther 2002; 40: 135-41.
- 22 Bijsterveld NR, Moons AH, Boekholdt SM. et al Ability of recombinant factor VIIa to reverse the anticoagulant effect of the pentasaccharide fondaparinux in healthy volunteers. Circulation 2002; 106: 2550-4.
- 23 Keularts IM, Hamulyak K, Hemker HC. et al The effect of DDAVP infusion on thrombin generation in platelet-rich plasma of von Willebrand type 1 and in mild haemophilia A patients. Thromb Haemost 2000; 84: 638-42.
- 24 Al Dieri R, Peyvandi F, Santagostino E. et al The thrombogram in rare inherited coagulation disorders: its relation to clinical bleeding. Thromb Haemost 2002; 88: 576-82.
- 25 Wielders S, Mukherjee M, Michiels J. et al The routine determination of the endogenous thrombin potential, first results in different forms of hyper-and hypocoagulability. Thromb Haemost 1997; 77: 629-36.
- 26 Kyrle PA, Mannhalter C, Béguin S. et al Clinical studies and thrombin generation in patients homozygous or heterozygous for the G20210A mutation in the prothrombin gene. Arterioscler Thromb Vasc Biol 1998; 18: 1287-91.
- 27 Hemker HC, Giesen PLA, Ramjee M. et al The thrombogram: monitoring thrombin generation in platelet rich plasma. Thromb Haemost 2000; 83: 589-91.
- 28 Ramjee MK. The use of fluorogenic substrates to monitor thrombin generation for the analysis of plasma and whole blood coagulation. Anal Biochem 2000; 277: 11-18.
- 29 Hemker HC, Willems GM, Béguin S. A computer assisted method to obtain the prothrom-bin activation velocity in whole plasma independent of thrombin decay processes. Thromb Haemost 1986; 56: 9-17.
- 30 Kessels H, Willems G, Hemker HC. Analysis of thrombin generation in plasma. Comput Biol Med 1994; 24: 277-88.
- 31 Sambrano GR, Weiss EJ, Zheng YW. et al Role of thrombin signaling in platelets in haemostasis and thrombosis. Nature 2001; 413: 74-8.
- 32 Carr Jr ME, Martin EJ, Carr SL. Delayed, reduced or inhibited thrombin production reduces platelet contractile force and results in weaker clot formation. Blood Coagul Fibrinolysis 2002; 13: 193-7.
- 33 Monroe DM, Roberts HR, Hoffman M. Platelet procoagulant complex assembly in a tissue factor-initiated system. Br J Haematol 1994; 88: 364-71.
- 34 Lawson JH, Kalafatis M, Stram S. et al A model for the tissue factor pathway to throm-bin. I. An empirical study. J Biol Chem 1994; 269: 23357-66.
- 35 Kjalke M, Ezban M, Monroe DM. et al High-dose factor VIIa increases initial thrombin generation and mediates faster platelet activation in thrombocytopenia-like conditions in a cell-based model system. Br J Haematol 2001; 114: 114-20.