Download PDF
Thromb Haemost 1986; 55(01): 074-077
DOI: 10.1055/s-0038-1661451
Original Article
Schattauer GmbH Stuttgart

A Sensitive Assay, Specific for Endothelial Cell Type Plasminogen Activator Inhibitor in Blood Plasma

E D Sprengers
The TNO Gaubius Institute for Cardiovascular Research, Leiden, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 16 August 1985

Accepted 07 November 1985

Publication Date:
19 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

A polyclonal antibody raised against plasminogen activator (PA-)inhibitor from endothelial cells fully precipitates the PA-inhibitor in endothelial cell conditioned medium but only a part of the PA-inhibitory activity in blood plasma. This indicates that the PA-inhibitory activity in blood plasma is not due to a single inhibitory component. Performing the assay for PA-inhibitory activity in plasma both in the presence and absence of saturating concentrations of anti-endothelial cell PA-inhibitor antibodies, allows the determination of endothelial cell type PA-inhibitor in plasma. The assay gives a linear dose-response curve of amount of plasma added versus t-PA neutralised.

Values for endothelial cell type PA-inhibitor in plasma of a group of 20 healthy individuals are in the range of 0.0-16.8 IU/ml and are not normally distributed (median value 3.0 IU/ml).

This method also reveals a second, so far unidentified, PA-inhibitory component in human plasma.

Key words

Fibrinolysis - Plasminogen activator inhibitor - Blood plasma
 

  • References

  • 1 Emeis JJ, Brommer EJ P, Kluft C, Brakman P. Progress in fibrinolysis. In Recent Adv Blood Coagulation Poller L. (ed) Churchill Livingstone, Edinburgh; 1985: 11-33
  • 2 Gaffney PJ. The fibrinolytic system. In Haemostasis and Thrombosis Bloom AC, Thomas DP. (Eds) Churchill Livingstone; New York: 1981: 198-224
  • 3 Collen D. On the regulation and control of fibrinolysis. Thromb Haemostas 1983; 43: 77-89
    PubMedGoogle Scholar
  • 4 Verheijen JH, Chang GT G, Kluft C. Evidence for the occurrence of a fast-acting inhibitor of plasminogen activator in human plasma. Thromb Haemostas 1984; 51: 392-395
    PubMedGoogle Scholar
  • 5 Kruithof EK O, Tran-Thang C, Ransijn A, Bachmann F. Demonstration of a fast-acting inhibitor of plasminogen activators in human plasma. Blood 1984; 64: 907-913
    PubMedGoogle Scholar
  • 6 Wiman B, Chmielewska I, Rånby M. Inactivation of tissue plasminogen activator in plasma. J Biol Chem 1984; 259: 3644-3647
    PubMedGoogle Scholar
  • 7 Thorsen S, Philips M. Isolation of tissue-type plasminogen activator-inhibitor complexes from human plasma. Biochim Biophys Acta 1984; 802: 111-118
    CrossrefPubMedGoogle Scholar
  • 8 Juhan-Vague I, Moerman B, DeCock F, Aillaud MF, Collen D. Plasma levels of a specific inhibitor of tissue-type plasminogen activator (and urokinase) in normal and pathological conditions. Thromb Res 1984; 33: 523-530
    CrossrefPubMedGoogle Scholar
  • 9 Korninger C, Wagner O, Binder BR. Tissue plasminogen activator inhibitor in human plasma: Development of a functional assay system and demonstration of a correlating Mr = 50000 antiactivator. J Lab Clin Med 1985; 105: 718-724
    PubMedGoogle Scholar
  • 10 Golder JP, Stephens RW. M inactivin: a human monocyte product which specifically inactivates urokinase-type plasminogen activators. Eur J Biochem 1983; 136: 517-522
    CrossrefPubMedGoogle Scholar
  • 11 Erickson LA, Ginsberg MH, Loskutoff DJ. Detection and partial characterisation of an inhibitor of plasminogen activator in human platelets. J Clin Invest 1984; 74: 1465-1472
    CrossrefPubMedGoogle Scholar
  • 12 Sprengers ED, Jansen BG. A fast-acting plasminogen activator inhibitor from human blood platelets. Thromb Haemostas 1985 54. 277 (Abstr)
    PubMedGoogle Scholar
  • 13 Holmberg L, Lecander I, Persson B, Astedt B. An inhibitor from placenta specifically binds urokinase and inhibits plasminogen activator released from ovarian carcinoma in tissue culture. Biochim Biophys Acta 1978; 544: 128-137
    CrossrefPubMedGoogle Scholar
  • 14 Wijngaards G. Interfering factors in the assay of plasminogen activators by the fibrin plate method. Thromb Haemostas 1979; 41: 590-600
    PubMedGoogle Scholar
  • 15 Lecander I, Roblin R, Åstedt B. Differential inhibition of two molecular forms of melanoma cell plasminogen activator by a placental inhibitor. Brit J Haematol 1984; 57: 407-412
    CrossrefPubMedGoogle Scholar
  • 16 Emeis JJ, Van Hinsbergh VW M, Verheijen JH, Wijngaards G. Inhibition of tissue-type plasminogen activator by conditioned medium from cultured human and porcine vascular endothelial cells. Biochem Biophys Res Comm 1983; 110: 392-398
    CrossrefPubMedGoogle Scholar
  • 17 Dosne AM, Dupuy E, Bodevin E. Production of a fibrinolytic inhibitor by cultured endothelial cells derived from human umbilical vein. Thromb Res 1978; 12: 377-387
    CrossrefPubMedGoogle Scholar
  • 18 Esnard F, Dupuy E, Dosne AM, Bodevin E. Partial characterization of a fibrinolytic inhibitor produced by cultured endothelial cells derived from human umbilical vein. Thromb Haemostas 1982; 47: 128-131
    PubMedGoogle Scholar
  • 19 Levin EG. Latent tissue plasminogen activator produced by human endothelial cells in culture: evidence for an enzyme-inhibitor complex. Proc Natl Acad Sci USA 1983; 80: 6804-6808
    CrossrefPubMedGoogle Scholar
  • 20 Philips M, Juul A, Thorsen S. Human endothelial cells produce a plasminogen activator inhibitor and a tissue-type plasminogen activator-inhibitor complex. Biochim Biophys Acta 1984; 802: 99-110
    CrossrefPubMedGoogle Scholar
  • 21 Loskutoff DJ, Van Mourik JA, Erickson LA, Lawrence D. Detection of an unusually stable fibrinolytic inhibitor produced by bovine endothelial cells. Proc Natl Acad Sci USA 1983; 80: 2956-2960
    CrossrefPubMedGoogle Scholar
  • 22 Sprengers ED, Verheijen JH, Van Hinsbergh VW M, Emeis JJ. Evidence for the presence of two different fibrinolytic inhibitors in human endothelial cell conditioned medium. Biochim Biophys Acta 1984; 801: 163-170
    CrossrefPubMedGoogle Scholar
  • 23 Van Mourik JA, Lawrence DA, Loskutoff DJ. Purification of an inhibitor of plasmingen activator (anti-activator) synthesized by endothelial cells. J Biol Chem 1984; 259: 14914-14921
    PubMedGoogle Scholar
  • 24 Sprengers ED, Princen HM G, Kooistra T, Van Hinsbergh VW M. Inhibition of plasminogen activators by conditioned medium of human hepatocytes and hepatoma cell line Hep G2. J Lab Clin Med 1985; 105: 751-758
    PubMedGoogle Scholar
  • 25 Sprengers ED, Van Hinsbergh VW M, Jansen BG. Relationship between the fast-acting and the latent plasminogen activator inhibitor, secreted by human endothelial cells. Thromb Haemostas 1985 54. 229 (Abstr.)
    PubMedGoogle Scholar
  • 26 Kluft C, Brakman P, Veldhuyzen-Stolk EC. Screening of fibrinolytic activity in plasma euglobulin fractions on the fibrin plate. In Progress in Chemical Fibrinolysis and Thrombolysis Davidson JF, Samama MM, Desnoyers PC. (Eds). Raven Press; New York: 1976. 2 37-65
  • 27 Kluft C, Van Wezel AL, Van der Velden CA M, Emeis JJ, Verheijen JH, Wijngaards G. Large-scale production of extrinsic (tissue-type) plasminogen activator from human melanoma cells. In Advances in Biotechnological Processes Mizrahi A, Van Wezel AL. (Eds). Alan R Liss; New York: 1983. 2 97-110
  • 28 Gaffney PJ, Curtis AD. A collaborative study of a proposed international standard for tissue-type plasminogen activator (t-PA). Thromb Haemostas 1985; 53: 134-136
    PubMedGoogle Scholar
  • 29 Verheijen JH, Mullaart E, Chang GT G, Kluft C, Wijngaards G. A simple, sensitive spectrophotometric assay for extrinsic (tissue-type) plasminogen activator applicable to measurements in plasma. Thromb Haemostas 1982; 48: 266-269
    PubMedGoogle Scholar