Reactivity of Soluble Fibrin Assays with Plasmic Degradation Products of Fibrin and in Patients Receiving Fibrinolytic Therapy

Bonnie I. McCarron; Victor J. Marder; Charles W. Francis

doi:10.1055/s-0037-1614905

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1999; 82(06): 1722-1729
DOI: 10.1055/s-0037-1614905

Rapid Communication

Schattauer GmbH

Reactivity of Soluble Fibrin Assays with Plasmic Degradation Products of Fibrin and in Patients Receiving Fibrinolytic Therapy

Bonnie I. McCarron

¹From the Vascular Medicine Unit, Department of Medicine, University of Rochester School of Medicine & Dentistry, Rochester, NY, USA

,

Victor J. Marder

¹From the Vascular Medicine Unit, Department of Medicine, University of Rochester School of Medicine & Dentistry, Rochester, NY, USA

,

Charles W. Francis

¹From the Vascular Medicine Unit, Department of Medicine, University of Rochester School of Medicine & Dentistry, Rochester, NY, USA

› Institutsangaben

Abstract

Summary

The ability to identify the products of thrombin and plasmin action on fibrinogen is important in patients with thrombotic and fibrinolytic disorders. New assays have been developed for “soluble fibrin” which represents soluble derivatives other than fibrinopeptides formed from fibrinogen by thrombin. These assays are either immunological, using antibodies for fibrin-specific neoepitopes, or functional and based on the cofactor activity of soluble fibrin in the tissue plasminogen activator (t-PA)-mediated conversion of plasminogen to plasmin. As plasmic derivations of fibrin share structural features with soluble fibrin, they may be reactive with assays for soluble fibrin. Therefore, we prepared plasmic digests of fibrin and determined the degree of reactivity with four soluble fibrin assays. Three assays used Mabs directed toward the fibrin-specific neoepitopes at α17-23 (A), γ312-324 (B) and α17-78 (D). A fourth (C) was based on t-PA co-factor activity. Tests A and C demonstrated marked crossreactivity with fibrin degradation products, and digests containing the largest derivatives showed greatest reactivity. Plasmic derivatives of crosslinked fibrin had greater reactivity than those of non-crosslinked fibrin. Tests B and D demonstrated minimal reactivity with plasmic derivatives of crosslinked or of non-crosslinked fibrin. Samples from patients with lower limb peripheral arterial occlusion were assayed for soluble fibrin, D-dimer and fibrinogen at presentation and eight hours after thrombolytic therapy. Variable results were seen at presentation with elevations in 13, 1, 0 and 4 of 19 patients using Tests A, B, C and D, respectively. After fibrinolytic therapy, marked increases in soluble fibrin levels were observed up to 600-fold above normal. A strong correlation between baseline levels was observed with Test B and Test D, which showed the least cross-reactivity with plasmic derivations. After thrombolytic therapy there were either weak or no correlations among the different assays. The results demonstrate that assays for soluble fibrin may react with plasmic derivatives of fibrin and this must be considered in interpreting clinical results.

Keywords

Soluble fibrin - fibrin(ogen) degradation products - fibrinolysis - peripheral vascular disease - fibrinolytic therapy - D-dimer

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References
1 Bauer KA, Rosenberg RD. The pathophysiology of the prethrombotic state in humans: Insights gained from studies using markers of hemostatic system activation. Blood 1987; 70: 343-50.
2 Teitel JM, Bauer KA, Lau HK, Rosenberg RD. Studies of the prothrombin activation pathway utilizing radioimmunoassays for the F₂/F₁₊₂ fragment and thrombin-antithrombin complex. Blood 1982; 59: 1086-97.
3 Bauer KA, Kass BL, ten Cate H, Bednarek MA, Hawiger JJ, Rosenberg RD. Detection of factor X activation in humans. Blood 1989; 74: 2007-15.
4 Lau HK, Rosenberg RD. The isolation and characterization of a specific antibody population directed against the thrombin antithrombin complex. J Biol Chem 1980; 225: 5885-93.
5 Nossel HL, Ti M, Kaplan KL, Spanondis K, Soland T, Butler Jr VP. The generation of fibrinopeptide A in clinical blood samples. Evidence for thrombin activity J Clin Invest 1976; 58: 1136-44.
6 Godal HC, Abildgaard U. Gelation of soluble fibrin in plasma by ethanol. Scand J Haemat 1966; 3: 342-50.
7 Niewiarowski S, Gurewich V. Laboratory identification of intravascular coagulation. The serial dilution protamine sulfate test for the detection of fibrin monomer and fibrin degradation products. J Lab Clin Med 1971; 77: 665-76.
8 Fletcher AP, Alkjaersig NK, Ghani FM, Tuleuski V, Owens O. Blood coagulation system pathophysiology in acute myocardial infarction: The influence of anticoagulant treatment on laboratory findings. J Lab Clin Med 1979; 93: 1054-65.
9 Largo R, Heller V, Straub PW. Detection of soluble intermediates of the fibrinogen-fibrin conversion using erythrocytes coated with fibrin monomers. Blood 1976; 47: 991-1001.
10 Connaghan DG, Francis CW, Lane DA, Marder VJ. Specific identification of fibrin polymers, fibrinogen degradation products, and crosslinked fibrin degradation products in plasma and serum with a new sensitive technique. Blood 1985; 65: 589-97.
11 Friberger P. Methods for determination of plasmin, antiplasmin and plasminogen by means of substrate S-2251. Haemostasis 1975; 7: 138-45.
12 McCarron BI, Marder VJ, Kanouse JJ, Francis CW. A soluble fibrin standard: Comparable dose-response with immunologic and functional assays. Report prepared on behalf of the DIC Subcommittee of the SSC. Thromb Haemost 1999; 82: 145-8.
13 Soe G, Kohno I, Inuzuka K, Itoh Y, Matsuda M. A monoclonal antibody that recognizes a neo-antigen exposed in the E domain of fibrin monomer complexed with fibrinogen or its derivatives: Its application to the measurement of soluble fibrin in plasma. Blood 1996; 88: 2109-17.
14 Francis CW, Marder VJ, Barlow GH. Plasmic degradation of crosslinked fibrin. Characterization of new macromolecular soluble complexes and a model of their structure. J Clin Invest 1980; 66: 1033-43.
15 Dempfle CE, Dollman M, Lill H, Puzzovio D, Dessauer A, Heene DL. Binding of a new monoclonal antibody against N-terminal heptapeptide of fibrin α-chain to fibrin polymerization site ‘A’: effects of fibrinogen and fibrinogen derivatives, and pretreatment of samples with NaSCN. Blood Coag Fibrinol 1993; 4: 79-86.
16 Lill H, Spannagl M, Trauner A, Schramm W, Schuller D, Ofenloch-Haehnle B, Draeger B, Naser W, Dessauer A. A new immunoassay for soluble fibrin enables a more sensitive detection of the activation state of blood coagulation in vivo. Blood Coag Fibrinol 1993; 4: 97-102.
17 Schielen WJG, Adams HPHM, van Leuven K, Voskuilen M, Tesser GI, Nieuwenhuizen W. The sequence γ-(312-324) is a fibrin-specific epitope. Blood 1991; 77: 2169-73.
18 Hoegee-de Nobel E, Voskuilen M, Briët E, Brommer EJP, Nieuwenhuizen W. A monoclonal antibody-based quantitative enzyme immunoassay for the determination of plasma fibrinogen concentrations. Thromb Haemost 1988; 60: 415-8.
19 Wiman B, Ranby M. Determination of soluble fibrin in plasma by rapid and quantitative spectrophotometric assay. Thromb Haemost 1986; 55: 189-93.
20 Olexa SA, Budzynski AZ. Evidence for four different polymerization sites involved in human fibrin formation. Proc Natl Acad Sci, USA 1980; 77: 1374-8.
21 Takagi T, Doolittle RF. Amino acid sequence studies on plasmin-derived fragments of human fibrinogen: Amino-terminal sequences of intermediate and terminal fragments. Biochem 1975; 24: 940-6.
22 Olexa SA, Budzynski AZ, Doolittle RF, Cottrell BA, Greene TC. Structure of fragment E species from human cross-linked fibrin. Biochem 1981; 20: 6139-45.
23 Nieuwenhuizen W. Sites in fibrin involved in the acceleration of plasminogen activation by t-PA. Possible role of fibrin polymerisation. Thromb Res 1994; 75: 343-7.
24 Marder VJ, Shulman NR, Carroll WR. High molecular weight derivatives of human fibrinogen produced by plasmin. I. Physicochemical and immunological characterization. J Biol Chem 1969; 244: 2111-9.
25 Bos R, Laterveer-Vreeswijk GH, Lockwood D, Szewczyk K, Nieuwenhuizen W. A new enzyme immunoassay for soluble fibrin in plasma, with a high discriminating power for thrombotic disorders. Thromb Haemost 1999; 81: 54-9.
26 Verheijen JH, Nieuwenhuizen W, Wijngaards G. Activation of plasminogen by tissue activator is increased specifically in the presence of certain soluble fibrin(ogen) fragments. Thromb Res 1982; 27: 377-85.
27 Bredbacka S, Blomback M, Wiman B, Pelzer H. Laboratory methods for detecting disseminated intravascular coagulation (DIC): new aspects. Acta Anaesth Scand 1993; 37: 125-30.
28 Wada H, Wakita Y, Nakase T, Shimura M, Hiyoyama K, Nagaya S, Deguchi H, Mori Y, Kaneko T, Deguchi K, Fujii J, Shiku H. Increased plasma-soluble fibrin monomer levels in patients with disseminated intravascular coagulation. Am J Hematol 1996; 51: 255-60.
29 Vogel G, Spanuth E. Predictive value of fibrin monomers in postoperative deep vein thrombosis. Klin Wochenschr 1990; 68: 1020-6.
30 Ginsberg JS, Siragusa S, Douketis J, Johnston M, Moffat K, Stevens P, Brill-Edwards P, Panju A, Patel A. Evaluation of a soluble fibrin assay in patients with suspected deep vein thrombosis. Thromb Haemost 1995; 74: 833-6.
31 Vogel G, Dempfle C-E, Spannagl M, Leskopf W. The value of quantitative fibrin monomer determination in the early diagnosis of postoperative deep vein thrombosis. Thromb Res 1996; 81: 241-51.
32 Ginsberg JS, Siragusa S, Douketis J, Johnston M, Moffat K, Donovan D, McGinnis J, Brill-Edwards P, Panju A, Patel A, Weitz J. Evaluation of a soluble fibrin assay in patients with suspected pulmonary embolism. Thromb Haemost 1996; 75: 551-4.
33 Brimble KS, Ginsberg JS. Evaluation of the combination of a bedside D-dimer assay and enzyme-linked immunosorbent soluble fibrin assay in patients with suspected venous thromboembolism. Thromb Res 1997; 88: 291-7.
34 Francis CW, Connaghan DG, Scott WL, Marder VJ. Increased plasma concentration of cross-linked fibrin polymers in acute myocardial infarction. Circulation 1987; 75: 1170-7.
35 Carville DGM, Dimitrijevic N, Walsh M, Digirolamo T, Brill EM, Drew N, Gargan PE. Thrombus precursor protein (TpP): marker of thrombosis early in the pathogenesis of myocardial infarction. Clin Chem 1996; 42: 1537-41.
36 Lee LV, Ewald GA, McKenzie CR, Eisenberg PR. The relationship of soluble fibrin and cross-linked fibrin degradation products to the clinical course of myocardial infarction. Arterioscler Thromb Vasc Biol 1997; 17: 628-33.
37 Terres W, Kümmel P, Sudrow A, Reuter H, Meinertz T, Hamm CW. Enhanced coagulation activation in troponin T-positive unstable angina pectoris. Am Heart J 1998; 135: 281-6.
38 Bremme K, Blombäck M. Hemostatic abnormalities may predict chronic hypertension after preeclampsia. Gynecol Obstet Invest 1996; 41: 20-6.
39 Östlund E, Bremme K, Wiman B. Soluble fibrin in plasma as a sign of activated coagulation in patients with pregnancy complications. Acta Obstet Gynecol Scand 1998; 77: 165-9.
40 Francis CW, Doughney K, Brenner B, Klingbiel K, Marder VJ. Increased immunoreactivity of plasma after fibrinolytic activation in an anti-DD ELISA system. Role of soluble crosslinked fibrin polymers. Circulation 1989; 79: 666-73.
41 Bloom AL. The release of thrombin from fibrin by fibrinolysin. Br J Haematol 1962; 8: 129-38.
42 Francis CW, Markham Jr RE, Barlow GH, Florack TM, Dobrzynski DM, Marder VJ. Thrombin activity of fibrin thrombi and soluble plasmic derivatives. J Lab Clin Med 1983; 102: 220-30.
43 Liu CY, Nossel HL, Kaplan KL. The binding of thrombin by fibrin. J Biol Chem 1979; 254: 10421-5.
44 Eisenberg PR, Sherman LA, Jaffe AS. Paradoxic elevation of fibrinopeptide A after streptokinase: Evidence for continued thrombosis despite intense fibrinolysis. J Am Coll Cardiol 1987; 10: 527-9.
45 Merlini PA, Bauer KA, Oltrona L, Ardissino D, Spinola A, Cattaneo M, Broccolino M, Mannucci PM, Rosenberg RD. Thrombin generation and activity during thrombolysis and concomitant heparin therapy in patients with acute myocardial infarction. J Am Coll Cardiol 1995; 25: 203-9.