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Thromb Haemost 2012; 108(05): 863-871
DOI: 10.1160/TH12-04-0269
Theme Issue Article
Schattauer GmbH

Coagulation on Endothelial Cells: The Underexposed Part of Virchow’s Triad

Irma L.A. Geenen
1   Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
2   Department of General Surgery, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
,
Mark J. Post
1   Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
,
Daniel G.M. Molin
1   Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
,
Geert W.H. Schurink
2   Department of General Surgery, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
,
Jos G. Maessen
3   Department of Cardiothoracic Surgery, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
,
Rene v. Oerle
4   Department of Internal Medicine, Laboratory of Clinical Thrombosis and Haemostasis, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
,
Hugo ten Cate
4   Department of Internal Medicine, Laboratory of Clinical Thrombosis and Haemostasis, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
,
Henri M.H. Spronk
4   Department of Internal Medicine, Laboratory of Clinical Thrombosis and Haemostasis, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center, Maastricht, The Netherlands
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Publikationsverlauf

Received: 29. April 2012

Accepted after major revision: 31. Juli 2012

Publikationsdatum:
29. November 2017 (online)

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Summary

The process of thrombin generation involves numerous plasma proteases and cofactors. Interaction with the vessel wall, in particular endothelial cells (ECs), influences this process but data on this interaction is limited. We evaluated thrombin generation on EA.hy926, human coronary arterial ECs (HCAECs) and patient-derived human venous ECs (HVECs) by means of a modified calibrated automated thrombogram (CAT) method and especially looked into contribution of the intrinsic and extrinsic pathways. Thrombin generation was measured in presence of confluent ECs with normal pooled and factor XII-deficient (FXII-deficient) platelet-poor plasma, with/without active site inhibited factor VIIa (ASIS) to block the extrinsic pathway and corn trypsin inhibitor for blocking contact activation (intrinsic pathway). Fetal bovine serum (FBS) was removed from culture conditions as FXIIa from the serum retained on ECs apparently, thereby inducing strong contact activation. In serum-free conditions, EA.hy926 and patient-derived HVECs induced thrombin generation mainly via the contact activation pathway with minor influence of ASIS on peak height and very low thrombin generation curves in FXII-deficient plasma. HVECs derived from coronary arterial bypass graft (CABG) patients showed increased thrombin generation compared to control patients, which could be ascribed to increased contact activation. Contribution of the extrinsic pathway on patient-derived ECs was limited. We conclude that the CAT method in combination with serum-free cultured ECs offers a valuable high-throughput method to evaluate endothelial influences on thrombin generation, which appears to involve predominantly contact activation on ECs. Contact activation-mediated thrombin generation was increased on ECs from CABG patients compared to controls.

Keywords

Endothelial cells - thrombin generation - cell culture - contact activation - extrinsic pathway
 

  • References

  • 1 Furie B, Furie BC. Mechanisms of Thrombus Formation. N Engl J Med 2008; 359: 938-949.
    CrossrefPubMedGoogle Scholar
  • 2 Bombeli T, Mueller M, Haeberli A. Anticoagulant properties of the vascular endothelium. Thromb Haemost 1997; 77: 408-423.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 3 Drake TA, Cheng J, Chang A. et al. Expression of tissue factor, thrombomodulin, and E-selectin in baboons with lethal Escherichia coli sepsis. Am J Pathol 1993; 142: 1458-1470.
    PubMedGoogle Scholar
  • 4 Kaikita K, Takeya M, Ogawa H. et al. Co-localization of tissue factor and tissue factor pathway inhibitor in coronary atherosclerosis. J Path 1999; 188: 180-188.
    CrossrefPubMedGoogle Scholar
  • 5 Wilcox JN, Smith KM, Schwartz SM. et al. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci USA 1989; 86: 2839-2843.
    CrossrefPubMedGoogle Scholar
  • 6 Lammle B, Wuillemin WA, Huber I. et al. Thromboembolism and bleeding tendency in congenital factor XII deficiency–a study on 74 subjects from 14 Swiss families. Thromb Haemost 1991; 65: 117-121.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 7 Renné T, Pozgajova M, Gruner S. et al. Defective thrombus formation in mice lacking coagulation factor XII. J Exp Med 2005; 202: 271-281.
    CrossrefPubMedGoogle Scholar
  • 8 Kleinschnitz C, Stoll G, Bendszus M. et al. Targeting coagulation factor XII provides protection from pathological thrombosis in cerebral ischemia without interfering with hemostasis. J Exp Med 2006; 203: 513-518.
    CrossrefPubMedGoogle Scholar
  • 9 Cheng Q, Tucker EI, Pine MS. et al. A role for factor XIIa, Äìmediated factor XI activation in thrombus formation in vivo. Blood 2010; 116: 3981-3989.
    CrossrefPubMedGoogle Scholar
  • 10 Hagedorn I, Schmidbauer S, Pleines I. et al. Factor XIIa Inhibitor Recombinant Human Albumin Infestin-4 Abolishes Occlusive Arterial Thrombus Formation Without Affecting Bleeding. Circulation 2010; 121: 1510-1517.
    CrossrefPubMedGoogle Scholar
  • 11 Smith SA, Mutch NJ, Baskar D. et al. Polyphosphate modulates blood coagulation and fibrinolysis. Proc Natl Acad Sci USA 2006; 103: 903-908.
    CrossrefPubMedGoogle Scholar
  • 12 van der Meijden PE, Munnix IC, Auger JM. et al. Dual role of collagen in factor XII-dependent thrombus formation. Blood 2009; 114: 881-890.
    CrossrefPubMedGoogle Scholar
  • 13 Schousboe I. Endothelial cells express a matrix protein which binds activated factor XII in a zinc-independent manner. Thromb Haemost 2006; 95: 312-319.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 14 Joseph K, Tholanikunnel BG, Ghebrehiwet B. et al. Interaction of high molecular weight kininogen binding proteins on endothelial cells. Thromb Haemost 2004; 91: 61-70.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 15 Campbell RA, Overmyer KA, Selzman CH. et al. Contributions of extravascular and intravascular cells to fibrin network formation, structure, and stability. Blood 2009; 114: 4886-4896.
    CrossrefPubMedGoogle Scholar
  • 16 Campbell JE, Brummel-Ziedins KE, Butenas S. et al. Cellular regulation of blood coagulation: a model for venous stasis. Blood 2010; 116: 6082-6091.
    CrossrefPubMedGoogle Scholar
  • 17 Roberts HR, Hoffman M, Monroe DM. A Cell-Based Model of Thrombin Generation. Semin Thromb Hemost 2006; 32 (Suppl. 01) 032 8.
    PubMedGoogle Scholar
  • 18 Edgell CJ, McDonald CC, Graham JB. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc Natl Acad Sci USA 1983; 80: 3734-3737.
    CrossrefPubMedGoogle Scholar
  • 19 Bouïs D, Hospers G, Meijer C. et al. Endothelium in vitro: A review of human vascular endothelial cell lines for blood vessel-related research. Angiogenesis 2001; 4: 91-102.
    CrossrefPubMedGoogle Scholar
  • 20 Loubele ST, Spek CA, Leenders P. et al. Activated protein C protects against myocardial ischemia/ reperfusion injury via inhibition of apoptosis and inflammation. Arterioscler Thromb Vasc Biol 2009; 29: 1087-1092.
    CrossrefPubMedGoogle Scholar
  • 21 Shariat-Madar Z, Mahdi F, Schmaier AH. Factor XI assembly and activation on human umbilical vein endothelial cells in culture. Thromb Haemost 2001; 85: 544-551.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 22 Virchow R. Gesammelte Abhandlungen zur wissenschaftlichen Medicin. Meidinger, Frankfurt a. M. 1856
    PubMedGoogle Scholar
  • 23 Hemker HC, Giesen P, AlDieri R. et al. The calibrated automated thrombogram (CAT): a universal routine test for hyper- and hypocoagulability. Pathophysiol Haemost Thromb 2002; 32: 249-253.
    CrossrefPubMedGoogle Scholar
  • 24 Spronk HM, Dielis AW, De Smedt E. et al. Assessment of thrombin generation II: Validation of the Calibrated Automated Thrombogram in platelet-poor plasma in a clinical laboratory. Thromb Haemost 2008; 100: 362-364.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 25 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.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 26 Hockin MF, Kalafatis M, Shatos M. et al. Protein C activation and factor Va inactivation on human umbilical vein endothelial cells. Arterioscler Thromb Vasc Biol 1997; 17: 2765-2775.
    CrossrefPubMedGoogle Scholar
  • 27 Bevilacqua MP, Pober JS, Majeau GR. et al. Recombinant tumor necrosis factor induces procoagulant activity in cultured human vascular endothelium: characterization and comparison with the actions of interleukin 1. Proc Natl Acad Sci USA 1986; 83: 4533-4537.
    CrossrefPubMedGoogle Scholar
  • 28 Mazzolai L, Silacci P, Bouzourene K. et al. Tissue factor activity is upregulated in human endothelial cells exposed to oscillatory shear stress. Thromb Haemost 2002; 87: 1062-1068.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 29 Nawroth PP, Stern DM. Modulation of endothelial cell hemostatic properties by tumor necrosis factor. J Exp Med 1986; 163: 740-745.
    CrossrefPubMedGoogle Scholar
  • 30 Campbell RA, Overmyer KA, Bagnell CR. et al. Cellular procoagulant activity dictates clot structure and stability as a function of distance from the cell surface. Arterioscler Thromb Vasc Biol 2008; 28: 2247-2254.
    CrossrefPubMedGoogle Scholar
  • 31 Schousboe I, Thomsen P, Van Deurs B. Factor XII binding to endothelial cells depends on caveolae. Eur J Biochem 2004; 271: 2998-3005.
    CrossrefPubMedGoogle Scholar
  • 32 Mahdi F, Madar ZS, Figueroa CD. et al. Factor XII interacts with the multiprotein assembly of urokinase plasminogen activator receptor, gC1qR, and cytokeratin 1 on endothelial cell membranes. Blood 2002; 99: 3585-3596.
    CrossrefPubMedGoogle Scholar
  • 33 Berrettini M, Schleef RR, Heeb MJ. et al. Assembly and expression of an intrinsic factor IX activator complex on the surface of cultured human endothelial cells. J Biol Chem 1992; 267: 19833-19839.
    PubMedGoogle Scholar
  • 34 Houston P, Dickson MC, Ludbrook V. et al. Fluid Shear Stress Induction of the Tissue Factor Promoter In Vitro and In Vivo Is Mediated by Egr-1. Arterioscler Thromb Vasc Biol 1999; 19: 281-289.
    CrossrefPubMedGoogle Scholar
  • 35 Lin MC, Almus-Jacobs F, Chen HH. et al. Shear stress induction of the tissue factor gene. J Clin Invest 1997; 99: 737-744.
    CrossrefPubMedGoogle Scholar
  • 36 Silverman MD, Manolopoulos VG, Unsworth BR. et al. Tissue factor expression is differentially modulated by cyclic mechanical strain in various human endothelial cells. Blood Coagul Fibrinolysis 1996; 7: 281-288.
    CrossrefPubMedGoogle Scholar
  • 37 Renné T, Gailani D. Role of Factor XII in hemostasis and thrombosis: clinical implications. Expert Rev Cardiovasc Ther 2007; 5: 733-741.
    CrossrefPubMedGoogle Scholar
  • 38 Borissoff JI, Heeneman S, Kilinc E. et al. Early atherosclerosis exhibits an enhanced procoagulant state. Circulation 2010; 122: 821-830.
    CrossrefPubMedGoogle Scholar
  • 39 Kilinç E, van Oerle R, Borissoff JI. et al. Factor XII Activation is Essential to Sustain the Procoagulant Effects of Particulate Matter. J Thromb Haemost 2011; 9: 1350-1367.
    CrossrefPubMedGoogle Scholar
  • 40 Ghebrehiwet B, CebadaMora C, Tantral L. et al. gC1qR/p33 Serves as a Molecular Bridge between the Complement and Contact Activation Systems and Is an Important Catalyst in Inflammation. In: Current Topics in Complement. USA: Springer; 2006: 95-105.
    Google Scholar
  • 41 Celermajer DS, Sorensen KE, Gooch VM. et al. Non-invasive detection of endothelial dysfunction in children and adults at risk of atherosclerosis. Lancet 1992; 340: 1111-1115.
    CrossrefPubMedGoogle Scholar
  • 42 Ross R. Atherosclerosis–an inflammatory disease. N Engl J Med 1999; 340: 115-126.
    CrossrefPubMedGoogle Scholar