Simultaneous Occurrence of Human Antibodies Directed against Fibrinogen, Thrombin, and Factor V Following Exposure to Bovine Thrombin: Effects on Blood Coagulation, Protein C Activation and Platelet Function

Vibhuti D Chouhan; Raul A De La Cadena; Chandrasekaran Nagaswami; John W Weisel; Mehdi Kajani; A Koneti Rao

doi:10.1055/s-0038-1655966

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1997; 77(02): 343-349
DOI: 10.1055/s-0038-1655966

Original Article

Schattauer GmbH Stuttgart

Simultaneous Occurrence of Human Antibodies Directed against Fibrinogen, Thrombin, and Factor V Following Exposure to Bovine Thrombin: Effects on Blood Coagulation, Protein C Activation and Platelet Function

Authors

Vibhuti D Chouhan

¹Sol Sherry Thrombosis Research Center, and Departments of Medicine and Pathology, Temple University School of Medicine, Philadelphia, PA, USA
Raul A De La Cadena

¹Sol Sherry Thrombosis Research Center, and Departments of Medicine and Pathology, Temple University School of Medicine, Philadelphia, PA, USA
Chandrasekaran Nagaswami

³Department of Cell and Development Biology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
John W Weisel

³Department of Cell and Development Biology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
Mehdi Kajani

²Albert Einstein Medical Center, Philadelphia, PA, USA
A Koneti Rao

¹Sol Sherry Thrombosis Research Center, and Departments of Medicine and Pathology, Temple University School of Medicine, Philadelphia, PA, USA

Abstract

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Summary

We describe a patient with severe epistaxis, prolonged coagulation tests and decreased plasma factor V following exposure to bovine topical thrombin. Patient IgG, but not normal IgG, showed binding to immobilized thrombin (bovine > human) and fibrinogen, and to factor V by Western blotting; the binding to thrombin was inhibited by hirudin fragment 54-65. Electron microscopy of rotary shadowed preparations showed complexes with IgG molecules attached near the ends of trinodular fibrinogen molecules. Patient IgG inhibited procoagulant, anticoagulant and cell-stimulating functions of thrombin demonstrated by inhibition of fibrinogen clotting, protein C activation and platelet aggregation; thrombin hydrolysis of S-2238 was not inhibited. The results suggest that the antibody is targeted against anion-binding exosite and not catalytic site of thrombin. Antifibrinogen antibodies have not been reported in patients exposed to bovine thrombin. There is a pressing need to re-evaluate the role of bovine thrombin as a therapeutic agent.

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References

References
1 Strieker RB, Lane PK, Leffert JD, Rodgers GM, Shuman MA, Corash L. Development of antithrombin antibodies following surgery in patient with prosthetic cardiac valves. Blood 1988; 72: 1375-1380
2 Flaherty MJ, Handerson R, Wener MH. Iatrogenic immunization with bovine thrombin:a mechanism of prolonged thrombin times after surgery. Ann Int Med 1989; 111: 631-634
3 Lawson JH, Pennell BJ, Olson JD, Mann KG. Isolation and characterization of an acquired antithrombin antibody. Blood 1990; 76: 2249-2257
4 Zehnder JL, Leung LLK. Development of antibodies to thrombin and factor V with recurrent bleeding in a patient exposed to topical bovine thrombin. Blood 1990; 76: 2011-2016
5 Sié P, Bezeaud A, Dupouy D, Archipoff G, Freyssinet JM, Dagoujon JM, Serre G, Guillin MC, Boneu B. An acquired antithrombin antibody directed toward the catalytic center of the enzyme. J Clin Invest 1991; 88: 290-296
6 Rapaport SI, Zivelin A, Minow RA, Hunter CS, Donnelly K. Clinical significance of antibodies to bovine and human thrombin and factor V after surgical use of bovine thrombin. Am J Clin Path 1992; 97: 84-91
7 Costa JM, Fiessinger JN, Capron L, Aiach M. Partial characterization of an autoantibody recognizing the secondary binding site(s) of thrombin in a patient with recurrent spontaneous arterial thrombosis. Thromb Haemost 1992; 67: 193-199
8 Nichols WL, Daniels TM, Fisher PK, Owen WG, Pineda AA, Mann KG. Antibodies to bovine thrombin and coagulation factor V associated with surgical use of topical bovine thrombin or fibrin “GLUE”:a frequent finding. Blood 1993; 82: 59a
9 Amaud E, Lafay M, Gaussem P, Picard V, Perrus MJ, Aiach M. An autoantibody directed against human thrombin anion binding exosite in a patient with arterial thrombosis:effects on platelets, endothelial cells and protein C activation. Blood 1994; 84: 1843-1850
10 LaSpada AR, Skalhegg BS, Henderson R, Schmer G, Pierce R, Chandler W. Brief report:Fatal hemorrhage in a patient with an acquired inhibitor of human thrombin. New Eng J Med 1995; 333: 494-497
11 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
12 De LaCadena RA, Wyshock EG, Kunapuli SP, Schultze RL, Miller M, Walz DA, Colman RW. Platelet thrombospondin interaction with human high and low molecular weight kininogens. Thromb Haemost 1994; 72: 125-131
13 Stubbs MT, Bode W. A player of many parts: the spot light falls on thrombin’s structure. Thromb Res 1993; 69: 01-58
14 Krstenansky JL, Mao SJT. Antithrombin properties of C-terminus of hirudin using synthetic unsulfated N^α-acetyl-hirudin_45-65 . FEBS Lett 1987; 211: 10-16
15 Veklich YI, Gorkun OV, Medved LV, Nieuwenhuizen W, Weisel JW. Carboxyl terminal portions of the α chains of fibrinogen and fibrin:localization by electron microscopy and the effects of isolated αc fragments on polymerization. J Biol Chem 1993; 268: 13577-13585
16 Suzuki S, Sakuragawa N. A study on the properties of commercial thrombin preparations. Thromb Res 1989; 53: 271-277
17 Doolittle RF. The structure and evolution of vertebrate fibrinogen. Ann NY Acad Sc 1984; 408: 13-26
18 Hoots WK, Carrell NA, Wagner RH, Cooper HA, Mc DonaghJ. A naturally occurring antibody that inhibits fibrin polymerization. New Eng J Med 1981; 304: 857-861
19 Galanakis DK, Ginzler EM, Fikrig SM. Monoclonal IgG anticoagulants delaying fibrin aggregation in two patients with systemic lupus erythematosus (SLE). Blood 1978; 52: 1037-1046
20 Marciniak E, Greenwood MF. Acquired coagulation inhibitor delaying fibrinopeptide release. Blood 1979; 53: 81-91
21 Ghosh S, McEvoy P, McVerry BA. Idiopathic autoantibody that inhibits fibrin monomer polymerization. Br J Haematol 1983; 53: 65-72
22 Pirkle H, Theodore I. Thrombin like enzymes from snake venoms. In: Handbook of Natural Toxins, vol 5. Tu AT. (ed). W. B. Saunders; Philadelphia, PA: 1991. pp 225-252
23 Fenton JW II. Thrombin specificity. Ann NY Acad Sci 1981; 405: 469-495
24 Butkowski RJ, Elion J, Downing MR, Mann KG. Primary structure of human prothrombin 2 and α-thrombin. J Biol Chem 1977; 252: 4942-4953
25 Vu TKH, Hung DT, Wheaton VI, Coughlin SR. Molecular cloning of functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 1991; 64: 1057-1068
26 Coughlin SR. Thrombin receptor structure and function. Thromb Haemost 1993; 70: 184-187