Effects of tissue factor, thrombomodulin and elevated clotting factor levels on thrombin generation in the calibrated automated thrombogram

 

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Summary

Elevated procoagulant levels have been correlated with increased thrombin generation in vitro and with increased venous thromboembolism (VTE) risk in epidemiological studies. hrombin generation tests are increasingly being employed as a high throughput method to provide a global measure of procoagulant activity in plasma samples. The objective of this study was to distinguish the effects of assay conditions [tissue factor (TF), thrombomodulin, platelets/lipids] and factor levels on thrombin generation parameters, and determine the conditions and parameters with the highest sensitivity and specificity for detecting elevated factor levels. Thrombin generation was measured using calibrated automated thrombography (CAT) in corn trypsin inhibitor (CTI)-treated platelet-free plasma (PFP) and plateletrich plasma (PRP). Statistical analysis was performed using logarithms of observed values with analysis of variance that accounted for experiment and treatment. he relative sensitivity of lag time (LT), time to peak (TTP), peak height and endogenous thrombin potential (ETP) to elevated factors XI, IX,VIII, X, and prothrombin was as follows: PFP initiated with 1 pM TF > PFP initiated with 5 pM TF > PRP initiated with 1 pM TF. For all conditions, inclusion of thrombomodulin prolonged the LT and decreased the peak and ETP; however, addition of thrombomodulin did not increase the ability of CAT to detect elevated levels of individual procoagulant factors. In conclusion, CAT conditions differentially affected the sensitivity of thrombin generation to elevated factor levels. Monitoring the peak height and/ or ETP following initiation of clotting in PFP with 1 pM TF was most likely to detect hypercoagulability due to increased procoagulant factor levels.

• References

• 1 Al RDieri, Peyvandi F, Santagostino E. et al. The thrombogram in rare inherited coagulation disorders: its relation to clinical bleeding. Thromb Haemost 2002; 88: 576-582.
  Article in Thieme ConnectPubMedGoogle Scholar
• 2 Butenas S, van Veer C, Mann KG. “Normal” thrombin generation. Blood 1999; 94: 2169-2178.
  PubMedGoogle Scholar
• 3 Regnault V, Beguin S, Lecompte T. Calibrated automated thrombin generation in frozen-thawed plateletrich plasma to detect hypercoagulability. Pathophysiol Haemost Thromb 2003; 33: 23-29.
  CrossrefPubMedGoogle Scholar
• 4 Siegemund A, Petros S, Siegemund T. et al. The endogenous thrombin potential and high levels of coagulation factor VIII, factor IX and factor XI. Blood Coagul Fibrinolysis 2004; 15: 241-244.
  CrossrefPubMedGoogle Scholar
• 5 Allen GA, Wolberg AS, Oliver JA. et al. Impact of procoagulant concentration on rate, peak and total thrombin generation in a model system. J Thromb Haemost 2004; 02: 402-413.
  CrossrefPubMedGoogle Scholar
• 6 Kraaijenhagen RA, Anker PS, Koopman MM. et al. High plasma concentration of factor VIIIc is a major risk factor for venous thromboembolism. Thromb Haemost 2000; 83: 5-9.
  Article in Thieme ConnectPubMedGoogle Scholar
• 7 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: 3698-3703.
  PubMedGoogle Scholar
• 8 Meijers JC, Tekelenburg WL, Bouma BN. et al. High levels of coagulation factor XI as a risk factor for venous thrombosis. N Engl J Med 2000; 342: 696-701.
  CrossrefPubMedGoogle Scholar
• 9 Kamphuisen PW, Rosendaal FR, Eikenboom JC. et al. Factor V antigen levels and venous thrombosis: risk profile, interaction with factor V leiden, and relation with factor VIII antigen levels. Arterioscler Thromb Vasc Biol 2000; 20: 1382-1386.
  CrossrefPubMedGoogle Scholar
• 10 van Hylckama AVlieg, van der Linden IK, Bertina RM. et al. High levels of factor IX increase the risk of venous thrombosis. Blood 2000; 95: 3678-3682.
  PubMedGoogle Scholar
• 11 Kyrle PA, Minar E, Hirschl M. et al. High plasma levels of factor VIII and the risk of recurrent venous thromboembolism. N Engl J Med 2000; 343: 457-462.
  CrossrefPubMedGoogle Scholar
• 12 de Visser MC, Poort SR, Vos HL. et al. Factor X levels, polymorphisms in the promoter region of factor X, and the risk of venous thrombosis. Thromb Haemost 2001; 85: 1011-1017.
  Article in Thieme ConnectPubMedGoogle Scholar
• 13 Hemker HC, Giesen P, Al RDieri. et al. Calibrated automated thrombin generation measurement in clotting plasma. Pathophysiol Haemost Thromb 2003; 33: 4-15.
  CrossrefPubMedGoogle Scholar
• 14 Chantarangkul V, Clerici M, Bressi C. et al. Thrombin generation assessed as endogenous thrombin potential in patients with hyperor hypo-coagulability. Haematologica 2003; 88: 547-554.
  PubMedGoogle Scholar
• 15 Dargaud Y, Beguin S, Lienhart A. et al. Evaluation of thrombin generating capacity in plasma from patients with haemophilia A and B. Thromb Haemost 2005; 93: 475-480.
  Article in Thieme ConnectPubMedGoogle Scholar
• 16 Duchemin J, Pan-Petesch B, Arnaud B. et al. Influence of coagulation factors and tissue factor concentration on the thrombin generation test in plasma. Thromb Haemost 2008; 99: 767-773.
  Article in Thieme ConnectPubMedGoogle Scholar
• 17 Keularts IM, Zivelin A, Seligsohn U. et al. The role of factor XI in thrombin generation induced by low concentrations of tissue factor. Thromb Haemost 2001; 85: 1060-1065.
  Article in Thieme ConnectPubMedGoogle Scholar
• 18 Hron G, Kollars M, Binder BR. et al. Identification of patients at low risk for recurrent venous thromboembolism by measuring thrombin generation. JAMA 2006; 296: 397-402.
  CrossrefPubMedGoogle Scholar
• 19 Tripodi A, Legnani C, Chantarangkul V. et al. High thrombin generation measured in the presence of thrombomodulin is associated with an increased risk of recurrent venous thromboembolism. J Thromb Haemost 2008; 06: 1327-1333.
  CrossrefPubMedGoogle Scholar
• 20 Besser M, Baglin C, Luddington R. et al. High rate of unprovoked recurrent venous thrombosis is associated with high thrombin-generating potential in a prospective cohort study. J Thromb Haemost 2008; 06: 1720-1725.
  CrossrefPubMedGoogle Scholar
• 21 Ten Cate-Hoek AJ, Dielis AW, Spronk HM. et al. Thrombin generation in patients after acute deep-vein thrombosis. Thromb Haemost 2008; 100: 240-245.
  Article in Thieme ConnectPubMedGoogle Scholar
• 22 Brandts A, van Hylckama AVlieg, Rosing J. et al. The risk of venous thrombosis associated with a high endogenous thrombin potential in the absence and presence of activated protein C. J Thromb Haemost 2007; 05: 416-418.
  CrossrefPubMedGoogle Scholar
• 23 Tripodi A, Martinelli I, Chantarangkul V. et al. The endogenous thrombin potential and the risk of venous thromboembolism. Thromb Res 2007; 121: 353-359.
  CrossrefPubMedGoogle Scholar
• 24 Dargaud Y, Trzeciak MC, Bordet JC. et al. Use of calibrated automated thrombinography +/− thrombomodulin to recognise the prothrombotic phenotype. Thromb Haemost 2006; 96: 562-567.
  Article in Thieme ConnectPubMedGoogle Scholar
• 25 van Hylckama AVlieg, Christiansen SC, Luddington R. et al. Elevated endogenous thrombin potential is associated with an increased risk of a first deep venous thrombosis but not with the risk of recurrence. Br J Haematol 2007; 138: 769-774.
  CrossrefPubMedGoogle Scholar
• 26 Gerotziafas GT, Depasse F, Busson J. et al. Towards a standardization of thrombin generation assessment: the influence of tissue factor, platelets and phospholipids concentration on the normal values of Thrombogram-Thrombinoscope assay. Thromb J 2005; 03: 16.
  CrossrefPubMedGoogle Scholar
• 27 Dargaud Y, Luddington R, Gray E. et al. Effect of standardization and normalization on imprecision of calibrated automated thrombography: an international multicentre study. Br J Haematol 2007; 139: 303-309.
  CrossrefPubMedGoogle Scholar
• 28 Luddington R, Baglin T. Clinical measurement of thrombin generation by calibrated automated thrombography requires contact factor inhibition. J Thromb Haemost 2004; 02: 1954-1959.
  CrossrefPubMedGoogle Scholar
• 29 Dargaud Y, Luddington R, Baglin TP. Elimination of contact factor activation improves measurement of platelet-dependent thrombin generation by calibrated automated thrombography at low-concentration tissue factor. J Thromb Haemost 2006; 04: 1160-1161.
  CrossrefPubMedGoogle Scholar
• 30 Vanschoonbeek K, Feijge MA, Van Kampen RJ. et al. Initiating and potentiating role of platelets in tissue factor-induced thrombin generation in the presence of plasma: subject-dependent variation in thrombogram characteristics. J Thromb Haemost 2004; 02: 476-484.
  CrossrefPubMedGoogle Scholar
• 31 Regnault V, Hemker HC, Wahl D. et al. Phenotyping the haemostatic system by thrombography--potential for the estimation of thrombotic risk. Thromb Res 2004; 114: 539-545.
  CrossrefPubMedGoogle Scholar
• 32 Dielis AW, Castoldi E, Spronk HM. et al. Coagulation factors and the protein C system as determinants of thrombin generation in a normal population. J Thromb Haemost 2008; 06: 125-131.
  PubMedGoogle Scholar
• 33 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.
  PubMedGoogle Scholar
• 34 Monkovic DD, Tracy PB. Functional characterization of human platelet-released factor V and its activation by factor Xa and thrombin. J Biol Chem 1990; 265: 17132-17140.
  PubMedGoogle Scholar
• 35 Bidot L, Jy W, Bidot Jr C. et al. Microparticle-mediated thrombin generation assay: increased activity in patients with recurrent thrombosis. J Thromb Haemost 2008; 06: 913-919.
  CrossrefPubMedGoogle Scholar
• 36 Ollivier V, Wang J, Manly D. et al. Detection of endogenous tissue factor levels in plasma using the calibrated automated thrombogram assay. Thromb Res. 2009 prepublished online April 3, 2009.
  PubMedGoogle Scholar
• 37 Tripodi A, Legnani C, Palareti G. et al. More on: high thrombin generation and the risk of recurrent venous thromboembolism. J Thromb Haemost 2009; 07: 906-907.
  CrossrefPubMedGoogle Scholar
• 38 Sauls DL, Banini AE, Boyd LC. et al. Elevated prothrombin level and shortened clotting times in subjects with type 2 diabetes. J Thromb Haemost 2007; 05: 638-639.
  CrossrefPubMedGoogle Scholar