Antithrombotic Effect of Human Recombinant Tissue Factor Pathway Inhibitor on Endotoxin-induced Intravascular Coagulation in Rats: Concerted Effect with Antithrombin

Summary

In our current study, we examined the antithrombotic effect of Chinese hamster ovary cell-derived human recombinant tissue factor pathway inhibitor (h-rTFPI) by intravenous injection of h-rTFPI with or without antithrombin into endotoxin-treated rats. An injection of h-rTFPI at a high dose (4 mg/kg of h-rTFPI or three doses of 1 mg/kg) significantly prevented the decrease of fibrinogen and factor VIII and the increase of fibrin/fibrinogen degradation products and glutamic-pyruvic transaminase in rats, while a single injection of 1 mg/kg of h-rTFPI or 250 U/kg of antithrombin did not significantly prevent intravascular coagulation. However, a simultaneous injection of 1 mg/kg of h-rTFPI and 250 U/kg of antithrombin did significantly prevent intravascular coagulation. From the studies on the clearance rate and immunohistochemical staining of injected h-rTFPI into normal rats, we found that most of the intravenously-injected h-rTFPI was localized on the central vein and sinusoids in the liver and catabolized via the proteoglycan-mediated pathway with a half-life of 48 min. These results suggest that h-rTFPI and antithrombin prevented endotoxin-induced intravascular coagulation in concert by binding to the vascular walls of the liver and by inhibiting fibrin formation on Kupffer cells in hepatic sinusoids.

• References

• 1 Edgington TS, Mackman N, Brand K, Ruf W. The structural biology of expression and function of tissue factor. Thromb Haemost 1991; 66: 67-79
• 2 Vinazzer H. Therapeutic use of antithrombin in shock and disseminated intravascular coagulation. Semin Thromb Hemost 1989; 15: 347-352
• 3 Wun TC. Tissue factor pathway inhibitor. In: Molecular Biology of Thrombosis and Hemostasis Roberts HR, High KA. (eds.). Marcel Dekker, Inc.; New York: 1995
• 4 Day KC, Hoffman LC, Palmier MO, Kretzmer KK, Huang MD, Pyla EY, Spokas E, Broze GJ jr, Warren TG, Wun TC. Recombinant lipoprotein-associated coagulation inhibitor inhibits tissue thromboplastin-induced intravascular coagulation in the rabbit. Blood 1990; 76: 1538-1545
• 5 Creasey AA, Chang ACK, Feigen L, Wun TC, Taylor FB jr, Hinshaw LB. Tissue factor pathway inhibitor reduces mortality from Escherichia coli septic shock. J Clin Invest 1993; 91: 2850-2860
• 6 Bregengard C, Nordfang O, Wildgoose P, Svendsen O, Hedner U, Diness V. The effect of two-domain tissue factor pathway inhibitor on endotoxin-induced disseminated intravascular coagulation in rabbits. Blood Coagul Fibrinol 1993; 4: 699-706
• 7 Holst J, Lindblad B, Bergqvist D, Nordfang O, Ostergaard PB, Petersen JGL, Nielsen G, Hedner U. Antithrombotic effect of recombinant truncated tissue factor pathway inhibitor (TFPI 1-161) in experimental venous thrombosis - a comparison with low molecular weight heparin. Thromb Haemost 1994; 71: 214-219
• 8 Carr C, Bild GS, Chang ACK, Peer GT, Palmier MO, Frazier RB, Gustafson ME, Wun TC, Creasey AA, Hinshaw LB, Taylor FB jr, Galluppi GR. Recombinant E. coli-derived tissue factor pathway inhibitor reduces coagu-lopathic and lethal effects in the baboon gram-negative model of septic shock. Circ Shock 1995; 44: 126-137
• 9 Palmier MO, Hall LJ, Reisch CM, Baldwin MK, Wilson AGE, Wun TC. Clearance of recombinant tissue factor pathway inhibitor (TFPI) in rabbits. Thromb Haemost 1992; 68: 33-36
• 10 Bregengaard C, Nordfang O, Ostergaard P, Petersen JGL, Meyn G, Diness V, Svendsen O, Hedner U. Pharmacokinetics of full length and two-domain tissue factor pathway inhibitor in combination with heparin in rabbits. Thromb Haemost 1993; 70: 454-457
• 11 Warshawsky I, Broze GJ jr, Schwarzt AL. The low density lipoprotein receptor-related protein mediates the cellular degradation of tissue factor pathway inhibitor. Proc Natl Acad Sci USA 1994; 91: 6664-6668
• 12 Warshawsky I, Bu G, Mast A, Saffitz JE, Broze GJ jr, Schwartz AL. The carboxy terminus of tissue factor pathway inhibitor is required for interacting with hepatoma cells in vitro and in vivo. J Clin Invest 1995; 95: 1773-1781
• 13 Narita M, Bu G, Olins GM, Higuchi DA, Herz J, Broze GJ jr, Schwartz AL. Two receptor systems are involved in the plasma clearance of tissue factor pathway inhibitor in vivo. J Biol Chem 1995; 270: 24800-24804
• 14 Enjyoji K, Miyata T, Kamikubo Y, Kato H. Effect of heparin on the inhibition of factor Xa by tissue factor pathway inhibitor: A segment Gly212-Phe243, of the third Kunitz domain is a heparin-binding site. Biochemistry 1995; 34: 5725-5735
• 15 Fraker PJ, Speck JC jr. Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a, 6a-diphenylglycol-uril. Biochem Biophys Res Commun 1978; 80: 849-857
• 16 Abumiya T, Enjyoji K, Kokawa T, Kamikubo Y, Kato H. An anti-tissue factor pathway inhibitor (TFPI) monoclonal antibody recognized the third Kunitz domain of free-form TFPI but not lipoprotein-associated forms in plasma. J Biochem 1995; 118: 178-182
• 17 Meszaros K, Aberle S, Dedrick R, Machovich R, Horwitz A, Birr C, Theofan G, Parent JB. Monocyte tissue factor induction by lipopolysaccha-ride (LPS): Dependence on LPS-binding protein and CD 14, and inhibition by a recombinant fragment of bactericidal/permeability-increasing protein. Blood 1994; 83: 2516-2525
• 18 Arai M, Mochida S, Ohno A, Ogata I, Obama H, Maruyama I, Fujiwara K. Blood coagulation equilibrium in rat liver microcirculation as evaluated by endothelial cell thrombomodulin and macrophage tissue factor. Thromb Res 1995; 80: 113-123
• 19 Moll T, Czyz M, Holzmuller H, Hofer-Warbinek R, Wagner E, Winkler H, Bach FH, Hofer E. Regulation of the tissue factor promoter in endothelial cells. Binding of NF-kB-, AP-1, and Spl-like transcription factors. J Biol Chem 1995; 270: 3849-3857
• 20 Nakahara Y, Miyata T, Hamuro T, Funatsu A, Miyagi M, Tsunasawa S, Kato H. Amino acid sequence and carbohydrate structure of a recombinant human tissue factor pathway inhibitor expressed in Chinese hamster ovary cells. One N- and two O-linked carbohydrate chains are located between Kunitz domains 2 and 3 and one N-linked is in Kunitz domain 2. Biochemistry 1996 in press
• 21 Werling RW, Zacharski LR, Kisiel W, Bajaj SP, Memoli VA, Rousseau SM. Distribution of tissue factor pathway inhibitor in normal and malignant human tissues. Thromb Haemost 1993; 69: 366-369
• 22 Takei A, Tashiro K, Nakagawa K, Enjyoji K, Kato H, Sueishi K. An immunohistochemical study on the localization of TFPI in tissues (Abstract in Japanese). Jpn J Thromb Haemost 1993; 4: 319
• 23 Enjyoji K, Kato H, Kamikubo Y. An interaction of TFPI with human endothelial cells (Abstract in Japanese). Seikagaku 1995; 67: 67