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DOI: 10.1160/TH05-10-0684
Hyperprothrombinaemia-induced APC resistance: Differential influence on fibrin formation and fibrinolysis
Financial support: Supported by a grant from Ministero della Università e della Ricerca Scientifica e Tecnologica (ex 60%).Publikationsverlauf
Received
19. Oktober 2005
Accepted after revision
25. Februar 2006
Publikationsdatum:
30. November 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/200604/lookinside/thumbnails/10-1160-th05-10-0684-1.jpg)
Summary
The prothrombin gene mutation G20210A is a common risk factor for thrombosis and has been reported to cause APC resistance. However, the inhibition of thrombin formation by APC not only limits fibrin formation but also stimulates fibrinolysis by reducing TAFI activation. We evaluated the influence of prothrombin G20210A mutation on the anticoagulant and fibrinolytic activities of APC (1 µg/ml). Thirtytwo heterozygous carriers and 32 non carriers were studied. APC anticoagulant activity was assessed by aPTT prolongation whereas APC fibrinolytic activity was determined by a microplate clot lysis assay. APC-induced aPTT prolongation was markedly less pronounced in carriers than in non carriers. On the contrary, fibrinolysis time was shortened by APC to a comparable extent in both groups. Accordingly, prothrombin levels were strongly correlated with APC-induced aPTT prolongation but not with APC-induced shortening of lysis time. The addition of purified prothrombin to normal plasma (final concentration 150%) caused APC resistance in the clotting assay over the whole range of tested APC concentrations (0.125–1.5 µg/ml). In the fibrinolytic assay, instead, prothrombin supplementation made the sample resistant to low but not to high concentrations of APC (>0.5 µg/ml). Thrombin and TAFIa determination in the presence of 1 µg/ml APC revealed that hyperprothrombinemia, although capable of enhancing thrombin generation, was unable to induce detectable TAFIa formation. It is suggested that APC resistance caused by hyperprothrombinaemia does not translate in impaired fibrinolysis, at least in the presence of high APC levels, because the increase in thrombin formation is insufficient to activate the amount of TAFI required to inhibit plasminogen conversion. These data might help to better understand the relationship between thrombin formation and fibrinolysis down-regulation.
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References
- 1 Vicente V, Gonzales-Conejero R, Rivera J. et al. The prothrombin gene variant 20210A in venous and arterial thrombosis. Haematologica 1999; 84: 356-62.
- 2 Poort SR, Rosendaal FR, Reitsma PH. et al. A common genetic variation in the 3’untranslated region of the prothrombin gene is associated with elevated plasma prothrombin levels and an increase in venous thrombosis. Blood 1996; 88: 3698-703.
- 3 Kyrle PA, Mannhalter C, Beguin 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.
- 4 Eikelboom JW, Ivey L, Ivey J. et al. Familial thrombophilia and the prothrombin 20210A mutation: association with increased thrombin generation and unusual thrombosis. Blood Coagul Fibrinolysis 1999; 10: 1-5.
- 5 Colucci M, Binetti BM, Tripodi A. et al. Hyperprothrombinemia associated with prothrombin G20210A mutation inhibits plasma fibrinolysis through a TAFImediated mechanism. Blood 2004; 103: 2157-61.
- 6 Leurs J, Hendriks D. CarboxypeptidaseU (TAFIa): a metallocarboxypeptidase with a distinct role in haemostasis and a possible risk factor for thrombotic disease. Thromb Haemost 2005; 94: 471-87.
- 7 Tripodi A, Chantarangkul V, Mannucci PM. Hyperprothrombinemia may result in acquired activated protein C resistance. Blood 2000; 96: 3295-6.
- 8 Curvers J, Thomassen MC, Rimmer J. et al. Effects of hereditary and acquired risk factors of venous thrombosis ona thrombin generation-based APC resistance test. Thromb Haemost 2002; 08: 5-11.
- 9 Smirnov MD, Safa O, Esmon NL. et al. Inhibition of activated protein C anticoagulant activity by prothrombin. Blood 1999; 94: 3839-46.
- 10 Bertina RM, van der Marel-van Nieuwkoop W, Loeliger EA. Spectrophotometric assays of prothrombin in plasma of patients using oral anticoagulants. Thromb Haemost 1979; 42: 1296-305.
- 11 Dahlback B, Carlsson M, Svensson PJ. Familial thrombophilia due to a previously unrecognized mechanism characterized by poor anticoagulant response to activated protein C: prediction of a cofactor to activated protein C. Proc Natl Acad Sci USA 1993; 90: 1004-8.
- 12 Colucci M, Pentimone A, Binetti BM. et al. Effect of heparin on TAFI-dependent inhibition of fibrinolysis: relative importance of TAFa generated by clot-bound and fluid phase thrombin. Thromb Haemost 2002; 88: 282-7.
- 13 Antovic JP, Schulman S, An SS. et al. Does an enzyme other than thrombin contribute to unexpected changes in the levels of the different forms of thrombin activatable fibrinolysis inhibitor in patients with hemophilia A, hemophilia B and von Willebrand disease?. Scand J Clin Lab Invest 2004; 64: 745-51.
- 14 Bouma BN, Meijers JCM. Thrombin activatable fibrinolysis inhibitor (TAFI, plasma procarboxypeptidase B, procarboxypeptidase R, procarboxypeptidase U). J Thromb Haemost 2003; 01: 1566-74.
- 15 Sakharov D, Plow EF, Rijken DC. On the mechanism of the antifibrinolytic activity of plasma carboxypeptidase B. J Biol Chem 1997; 272: 14477-82.
- 16 Bajzar L, Nesheim ME, Tracy PB. The profibrinolytic effect of activated protein C in clots formed from plasma is TAFI-dependent. Blood 1996; 88: 2093-100.
- 17 Esmon CT. The regulation of natural anticoagulant pathways. Science 1987; 235: 1348-52.
- 18 Colucci M, Simioni P, Piro D. et al. An antifibrinolytic effect associated with an anti-factor V antibody in a patient with severe thrombophilia. Haematologica 2003; 88: 1383-9.
- 19 Bajzar L, Kalafatis M, Simioni P. et al. An antifibrinolytic mechanism describing the prothrombotic effect associated with factor VLeiden . J Biol Chem 1996; 271: 22949-52.
- 20 Frere C, Morange PE, Saut N. et al. Quantification of thrombin activatable fibrinolysis inhibitor (TAFI) gene polymorphism effects on plasma levels of TAFI measured with assays insensitive to isoform-dependent artefact. Thromb Haemost 2005; 94: 373-9.
- 21 Sakata Y, Curriden S, Lawrence D. et al. Activated protein C stimulates the fibrinolytic activity of cultured endothelial cells and decreases antiactivator activity. Proc Natl Acad Sci USA 1985; 82: 1121-5.
- 22 de Fouw NJ, van Tilburg NH, Haverkate F. et al. Activated protein C accelerates clot lysis by virtue of its anticoagulant activity. Blood Coagul Fibrinolysis 1993; 04: 201-10.
- 23 Broze Jr GJ. Thrombin-dependent inhibition of fibrinolysis. Curr Opin Hematol 1996; 03: 390-4.
- 24 Von dem Borne PA, Bajzar L, Meijers JC. et al. Thrombin-mediated activation of factor XI results in a thrombin-activatable fibrinolysis inhibitor-dependent inhibition of fibrinolysis. J Clin Invest 1997; 99: 2323-7.
- 25 Leurs J, Nerme V, Sim Y. et al. Carboxypeptidase U (TAFIa) prevents lysis from proceeding into the propagation phase through a threshold-dependent mechanism. J Thromb Haemost 2004; 02: 416-23.
- 26 Walker JB, Bajzar L. The intrinsic threshold of the fibrinolytic system is modulated by basic carboxypeptidases, but the magnitude of the antifibrinolytic effect of activated thrombin-activable fibrinolysis inhibitor is masked by its instability. J Biol Chem 2004; 279: 27896-904.
- 27 Esmon CT. Inflammation and thrombosis. J Thromb Haemost 2003; 01: 1343-8.
- 28 Comp PC, Jacocks RM, Ferrell GL. et al. Activation of protein C in vivo . J Clin Invest 1982; 70: 127-34.
- 29 Colucci M, Triggiani R, Cavallo LG. et al. Thrombin infusion in endotoxin-treated rabbits reduces the plasma levels of plasminogen activator inhibitor: evidence for a protein-C-mediated mechanism. Blood 1989; 74: 1976-82.
- 30 Vossen CY, Strandberg K, Stenflo J. et al. No effect of the prothrombin G20210A mutation on protein C activation in a large kindred with type I protein C deficiency. Blood Coagul Fibrinolysis 2004; 15: 573-6.
- 31 Mosnier LO, Meijers JC, Bouma BN. Regulation of fibrinolysis in plasma by TAFI and protein C is dependent on the concentration of thrombomodulin. Thromb Haemost 2001; 85: 5-11.