The Thrombin Activation Pathway Modulates the Assembly, Structure and Lysis of Human Plasma Clots In Vitro

Summary

Thrombin activation of the soluble plasma protein fibrinogen is vital for successful haemostasis. Thrombin is generated from prothrombin by the prothrombinase complex which also includes factor Xa, factor Va, Ca²⁺ and a procoagulant membrane surface. Factor X activation is catalysed in a complex including either factor Vila and tissue factor, or factor IXa and factor Villa. Factor IXa can be generated either by the factor Vlla/tissue factor complex or by factor XIa which is in turn produced by the contact phase reactions in vitro. Once activated, fibrinogen develops into the fibrin polymeric matrix at the site of injury. It is not known to what extent the properties of this haemostatic plug are sensitive to the pathway leading up to thrombin generation. Here static human plasma is studied in vitro using magnetically induced birefringence. It is shown that the contact phase/factor XIa pathway gives rise to linear fibrin assembly progress curves whereas the factor Vlla/tissue factor activation of factor X provokes largely sigmoid assembly. The latter pathway also causes the formation of significantly thicker fibres even although assembly is more rapid. This result is the inverse of that anticipated from the study of simple model systems. Whilst the streptokinase activated lysis of both types of clot exhibits similar biphasic kinetics, an exponential main phase followed by a sigmoidal tailing off, the data suggest that clots produced by the contact phase/factor XIa pathway are more recalcitrant to lysis. These results demonstrate that the profile of thrombin generation not only determines the kinetics of assembly but also influences the rate of lysis and structure of the haemostatic plug.

• References

• 1 Davie EW, Fujikawa K. Kisiel W The coagulation cascade: initiation, maintenance, and regulation. Biochemistry 1991; 30: 10363-70
• 2 Bolton-Maggs PH B, Young-Wan-Yin R, McCraw A, Slack J, Kemof PB A. Inheritance and bleeding in factor XI deficiency. Br J Haematol 1988; 69: 521-528
• 3 Ragni MV, Sinha D, Seaman F, Lewis JH, Spero JA, Walsh PN. Comparison of bleeding tendency, factor XI coagulant activity, and factor XI antigen in 25 factor XI-deficient kindreds. Blood 1985; 65: 719-724
• 4 Gailani D, Broze Jr GJ. Factor XI activation in a revised model of blood coagulation. Science 1991; 253: 909-912
• 5 Naito K, Fujikawa K. Activation of human blood coagulation factor XI independent of factor XII: Factor XI is activated by thrombin and factor XIa in the presence of negatively charged surfaces. J Biol Chem 1991; 266: 7353-7358
• 6 Scott CF, Colman RW. Fibrinogen blocks the autoactivation and thrombinmediated activation of factor XI on dextran sulfate. Proc Natl Acad Sci USA 1992; 89: 11189-11193
• 7 Gailani D, Broze GJ. Factor XII-independent activation of factor XI in plasma: effects of sulfatides on tissue factor-induced coagulation. Blood 1993; 82: 813-819
• 8 Lawson HJ, Kalafatis M, Stram S, Mann KG. An empirical study. J Biol Chem 1994; 269: 23357-23366
• 9 Nemerson Y. Tissue factor and hemostasis. Blood 1988; 71: 1-8
• 10 Østerud B, Rapaport SI. Activation of factor IX by the reaction product of tissue factor and factor VII. Additional pathway for initiating blood coagulation Proc Natl Acad Sci USA 1977; 74: 5260-5264
• 11 Komiyama Y, Pedersen AH, Kisiel W. Proteolytic activation of human factors IX and X by recombinant human factor VIIa: effects of calcium, phospholipids, and tissue factor. Biochemistry 1990; 29: 9418-9425
• 12 Ferry JD, Morrison PR. Preparation and properties of serum and plasma proteins VIII. Conversion of human fibrinogen to fibrin under various conditions J Am Chem Soc 1947; 69: 388-400
• 13 Blombäck B, Hessel B, Hogg D, Therkildsen L. A two-step fibrinogenfibrin transition in blood coagulation. Nature 1987; 275: 501-505
• 14 Hantgan RR, Hermans J. Assembly of Fibrin. J Biol Chem 1979; 254: 11272-11281
• 15 Weisel JW, Nagaswami C. Computer modeling of fibrin polymerization kinetics correlated with electron microscope and turbidity observations: clot structure and assembly are kinetically controlled. Biophys J 1992; 63: 111-128
• 16 Binnie CG, Lord ST. The fibrinogen sequences that interact with thrombin. Blood 1993; 81: 3186-3192
• 17 Bänninger H, Lämmle B, Furan M. Binding of α-thrombin to fibrin depends on the quality of the fibrin network. Biochem. J 1994; 298: 157-163
• 18 Greenberg CS, Shuman MA. The zymogen forms of blood coagulation factor XIII bind specifically to fibrinogen. J Biol Chem 1982; 257: 6096-6101
• 19 Mosher D. Cross-linking of cold insoluble globulin by fibrin stabilizing factor. J Biol Chem 1975; 250: 6614-6621
• 20 Bale MD, Mosher DF. Effects of thrombospondin on fibrin polymerisation and structure. J Biol Chem 1986; 261: 862-868
• 21 Entwistle RA, Furcht LT. C1q component of complement binds to fibrinogen and fibrin. Biochemistry 1988; 27: 507-512
• 22 Garman AM, Smith RA. The binding of plasminogen to fibrin: evidence for plasminogen-bridging. Throm Res 1982; 27: 311-320
• 23 Petersen LC, Suenson E. Effect of plasminogen and tissue-type plasminogen activator on fibrin gel structure. Fibrinolysis 1991; 5: 51-59
• 24 Reilly CF, Hutzelmann JE. Plasminogen activator inhibitor-1 binds to fibrin and inhibits tissue-type plasminogen activator-mediated fibrin dissolution. J Biol Chem 1992; 267: 17128-17135
• 25 Sakata Y, Aoki N. Cross-linking of α₂-plasmin inhibitor to fibrin by fibrin stabilizing factor. J Clin Inves 1980; 65: 290-297
• 26 Liu J, Harpel PC, Gurewich V. Fibrin-bound lipoprotein (a) promotes plasminogen binding but inhibits fibrin degradation by plasmin. Biochemistry 1994; 33: 2554-2560
• 27 Torbet J. Fibrin assembly in human plasma and fibrinogen/albumin mixtures. Biochemistry 1986; 25: 5309-5314
• 28 Carr ME. Fibrin formed in plasma is composed of fibres more massive than those formed from purified fibrinogen. Thromb Haemost 1988; 59: 535-539
• 29 Shah GA, Nair CH, Dhall DP. Comparison of fibrin networks in plasma and fibrinogen solution. Thromb Res 1987; 45: 257-264
• 30 Freyssinet JM, Torbet J, Hudry-Clergeon G, Maret G. Fibrinogen and fibrin structure and fibrin formation using magnetic orientation. Proc Natl Acad Sci USA 1983; 80: 1616-1620
• 31 Clauss A. Gerinnungsphysiologische Schnellmethode zur Bestimmung des Fibrinogen. Acta Haematol 1957; 17: 237-246
• 32 Hoffmann JJ M L, Vijgen M, Nieuwenhuizen W. Comparison of the specificity of fibrinogen assays during thrombolytic therapy. Fibrinolysis 1990; 4 Supp (Suppl. 02) 121-123
• 33 Maret G, Weill G. Magnetic birefringence study of the electrostatic and intrinsic persistence length of DNA. Biopolymers 1983; 22: 2727-2744
• 34 Torbet J, Freyssinet JM, Hudry-Clergeon G. Oriented fibrin gels formed by polymerisation in a strong magnetic field. Nature 1981; 289: 91-93
• 35 Freyssinet JM, Torbet J, Hudry-Clergeon G. Fibrinogen and fibrin in strong magnetic fields. Complementary results and discussion Biochimie 1984; 66: 81-85
• 36 Bishop MF, Ferrone FA. Kinetics of nucleation-controlled polymerization. A perturbation treatment for use with a secondary pathway Biophys J 1984; 46: 631-644
• 37 Torbet J. Anisotropy in spherical viruses demonstrated with magnetic birefringence. EMBO J 1983; 2: 63-66
• 38 Torbet J. Magnetic Birefringence Study of Fibrin Formation & Chylomicron Behaviour in Human Blood Plasma. In: Biophysical Effects of Steady Magnetic Fields. Maret G, Kiepenheuer J, Boccara N. eds Springer Verlag; 1986: 23-27
• 39 Collen D, Stump DC, Gold HK. Thrombolytic therapy. Ann Rev Med 1988; 39: 405-423
• 40 Iwasaka M, Ueno S, Tsuda H. Effects of magnetic fields on fibrinolysis. J Appl Phys 1994; 75: 7162-7164
• 41 Gabriel DA, Muga K, Boothroyd EM. The effect of fibrin structure on fibrinolysis. J Biol Chem 1992; 267: 24259-24263
• 42 Carrell N, Gabriel DA, Blatt PM, Carr ME, McDonagh J. Hereditary dysfi-brinogenemia in a patient with thrombotic disease. Blood 1983; 62: 439-447
• 43 Sakata Y, Mimuro J, Aoki N. Differential binding of plasminogen to cross- linked and noncrosslinked fibrins: Its significance in hemostatic defect in factor XIII deficiency. Blood 1984; 63: 1393-1401
• 44 Suenson E, Lützen O, Thorsen S. Initial plasmin-degradation of fibrin as the basis of a positive feed-back mechanism in fibrinolysis. Eur J Biochem 1984; 140: 513-522
• 45 Wun T-C, Capuano A. Spontaneous fibrinolysis in whole human plasma: Identification of tissue activator-related protein as the major plasminogen activator causing spontaneous activity in vitro. J Biol Chem 1985; 260: 5061-5066
• 46 Repke D, Gemmell CH, Guha A, Turitto VT, Broze GT, Nemerson Y. Hemophilia as a defect of the tissue factor pathway of blood coagulation: Effect of factors VIII and IX on factor X activation in a continuous-flow reaction. Proc Natl Acad Sci USA 1990; 87: 7623-7627
• 47 Torbet J, Ronziere MC. Magnetic alignment of collagen during self assembly. Biochem J 1984; 219: 1057-1059
• 48 Guido S, Tranquillo RT. A method for the systematic and quantitative study of cell contact guidance in oriented collagen gels: Correlation of fibroblast orientation and gel birefringence. J Cell Sci 1993; 105: 317-331