Skip to main content
NCBI home page
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1993 Oct;46(10):908–911. doi: 10.1136/jcp.46.10.908

Role of beta 2-glycoprotein I and anti-phospholipid antibodies in activation of protein C in vitro.

D M Keeling 1, A J Wilson 1, I J Mackie 1, D A Isenberg 1, S J Machin 1
PMCID: PMC501616  PMID: 8227406

Abstract

AIMS--To investigate the effect of beta 2-glycoprotein I (beta 2 GPI) on the thrombin/thrombomodulin dependent activation of protein C; and to determine whether beta 2 GPI dependent anticardiolipin antibodies have any effect. METHODS--Protein C was activated by thrombin in the presence of thrombomodulin and phospholipid vesicles in an in vitro system. The effect of adding purified beta 2 GPI to this system was observed. Affinity purified anticardiolipin antibodies and total IgG from patients with anticardiolipin antibodies and the lupus anticoagulant were studied for their effects on protein C activation in the presence and absence of beta 2 GPI. RESULTS--beta 2-Glycoprotein I had no effect on the activity of preformed activated protein C. When the phospholipid vesicles were incubated with beta 2 GPI before the addition of protein C, the activation of protein C was inhibited in a dose dependent manner. With phosphatidylserine:phosphatidylcholine vesicles at a concentration of 1 microM:2 microM, beta 2 GPI began to inhibit the reaction at a concentration of 15 nM, and at 4 microM (the normal plasma concentration) the activation of protein C was reduced to 40%. Anticardiolipin antibodies had no demonstrable effect. CONCLUSIONS--beta 2-Glycoprotein I inhibits protein C activation in an in vitro system. Its physiological role is unknown but it has potential procoagulant as well as anticoagulant properties. An effect of antiphospholipid antibodies on protein C activation, which might explain their association with thrombosis, could not be shown.

Full text

PDF
908

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BOWIE E. J., THOMPSON J. H., Jr, PASCUZZI C. A., OWEN C. A., Jr THROMBOSIS IN SYSTEMIC LUPUS ERYTHEMATOSUS DESPITE CIRCULATING ANTICOAGULANTS. J Lab Clin Med. 1963 Sep;62:416–430. [PubMed] [Google Scholar]
  2. Bancsi L. F., van der Linden I. K., Bertina R. M. Beta 2-glycoprotein I deficiency and the risk of thrombosis. Thromb Haemost. 1992 Jun 1;67(6):649–653. [PubMed] [Google Scholar]
  3. Bevers E. M., Galli M., Barbui T., Comfurius P., Zwaal R. F. Lupus anticoagulant IgG's (LA) are not directed to phospholipids only, but to a complex of lipid-bound human prothrombin. Thromb Haemost. 1991 Dec 2;66(6):629–632. [PubMed] [Google Scholar]
  4. Boey M. L., Colaco C. B., Gharavi A. E., Elkon K. B., Loizou S., Hughes G. R. Thrombosis in systemic lupus erythematosus: striking association with the presence of circulating lupus anticoagulant. Br Med J (Clin Res Ed) 1983 Oct 8;287(6398):1021–1023. doi: 10.1136/bmj.287.6398.1021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Borrell M., Sala N., de Castellarnau C., Lopez S., Gari M., Fontcuberta J. Immunoglobulin fractions isolated from patients with antiphospholipid antibodies prevent the inactivation of factor Va by activated protein C on human endothelial cells. Thromb Haemost. 1992 Sep 7;68(3):268–272. [PubMed] [Google Scholar]
  6. Canfield W. M., Kisiel W. Evidence of normal functional levels of activated protein C inhibitor in combined Factor V/VIII deficiency disease. J Clin Invest. 1982 Dec;70(6):1260–1272. doi: 10.1172/JCI110725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cariou R., Tobelem G., Bellucci S., Soria J., Soria C., Maclouf J., Caen J. Effect of lupus anticoagulant on antithrombogenic properties of endothelial cells--inhibition of thrombomodulin-dependent protein C activation. Thromb Haemost. 1988 Aug 30;60(1):54–58. [PubMed] [Google Scholar]
  8. De Wolf F., Carreras L. O., Moerman P., Vermylen J., Van Assche A., Renaer M. Decidual vasculopathy and extensive placental infarction in a patient with repeated thromboembolic accidents, recurrent fetal loss, and a lupus anticoagulant. Am J Obstet Gynecol. 1982 Apr 1;142(7):829–834. doi: 10.1016/s0002-9378(16)32527-3. [DOI] [PubMed] [Google Scholar]
  9. Elias M., Eldor A. Thromboembolism in patients with the 'lupus'-type circulating anticoagulant. Arch Intern Med. 1984 Mar;144(3):510–515. [PubMed] [Google Scholar]
  10. Firkin B. G., Howard M. A., Radford N. Possible relationship between lupus inhibitor and recurrent abortion in young women. Lancet. 1980 Aug 16;2(8190):366–366. doi: 10.1016/s0140-6736(80)90361-x. [DOI] [PubMed] [Google Scholar]
  11. Freyssinet J. M., Gauchy J., Cazenave J. P. The effect of phospholipids on the activation of protein C by the human thrombin-thrombomodulin complex. Biochem J. 1986 Aug 15;238(1):151–157. doi: 10.1042/bj2380151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Freyssinet J. M., Wiesel M. L., Gauchy J., Boneu B., Cazenave J. P. An IgM lupus anticoagulant that neutralizes the enhancing effect of phospholipid on purified endothelial thrombomodulin activity--a mechanism for thrombosis. Thromb Haemost. 1986 Jun 30;55(3):309–313. [PubMed] [Google Scholar]
  13. Galli M., Comfurius P., Barbui T., Zwaal R. F., Bevers E. M. Anticoagulant activity of beta 2-glycoprotein I is potentiated by a distinct subgroup of anticardiolipin antibodies. Thromb Haemost. 1992 Sep 7;68(3):297–300. [PubMed] [Google Scholar]
  14. Galli M., Comfurius P., Maassen C., Hemker H. C., de Baets M. H., van Breda-Vriesman P. J., Barbui T., Zwaal R. F., Bevers E. M. Anticardiolipin antibodies (ACA) directed not to cardiolipin but to a plasma protein cofactor. Lancet. 1990 Jun 30;335(8705):1544–1547. doi: 10.1016/0140-6736(90)91374-j. [DOI] [PubMed] [Google Scholar]
  15. Gharavi A. E., Harris E. N., Asherson R. A., Hughes G. R. Anticardiolipin antibodies: isotype distribution and phospholipid specificity. Ann Rheum Dis. 1987 Jan;46(1):1–6. doi: 10.1136/ard.46.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harris E. N., Gharavi A. E., Patel S. P., Hughes G. R. Evaluation of the anti-cardiolipin antibody test: report of an international workshop held 4 April 1986. Clin Exp Immunol. 1987 Apr;68(1):215–222. [PMC free article] [PubMed] [Google Scholar]
  17. Henry M. L., Everson B., Ratnoff O. D. Inhibition of the activation of Hageman factor (factor XII) by beta 2-glycoprotein I. J Lab Clin Med. 1988 May;111(5):519–523. [PubMed] [Google Scholar]
  18. Keeling D. M., Wilson A. J., Mackie I. J., Isenberg D. A., Machin S. J. Lupus anticoagulant activity of some antiphospholipid antibodies against phospholipid bound beta 2 glycoprotein I. J Clin Pathol. 1993 Jul;46(7):665–667. doi: 10.1136/jcp.46.7.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Keeling D. M., Wilson A. J., Mackie I. J., Machin S. J., Isenberg D. A. Some 'antiphospholipid antibodies' bind to beta 2-glycoprotein I in the absence of phospholipid. Br J Haematol. 1992 Nov;82(3):571–574. doi: 10.1111/j.1365-2141.1992.tb06469.x. [DOI] [PubMed] [Google Scholar]
  20. Malia R. G., Kitchen S., Greaves M., Preston F. E. Inhibition of activated protein C and its cofactor protein S by antiphospholipid antibodies. Br J Haematol. 1990 Sep;76(1):101–107. doi: 10.1111/j.1365-2141.1990.tb07843.x. [DOI] [PubMed] [Google Scholar]
  21. Marciniak E., Romond E. H. Impaired catalytic function of activated protein C: a new in vitro manifestation of lupus anticoagulant. Blood. 1989 Nov 15;74(7):2426–2432. [PubMed] [Google Scholar]
  22. Matsuura E., Igarashi Y., Fujimoto M., Ichikawa K., Koike T. Anticardiolipin cofactor(s) and differential diagnosis of autoimmune disease. Lancet. 1990 Jul 21;336(8708):177–178. doi: 10.1016/0140-6736(90)91697-9. [DOI] [PubMed] [Google Scholar]
  23. McNeil H. P., Simpson R. J., Chesterman C. N., Krilis S. A. Anti-phospholipid antibodies are directed against a complex antigen that includes a lipid-binding inhibitor of coagulation: beta 2-glycoprotein I (apolipoprotein H). Proc Natl Acad Sci U S A. 1990 Jun;87(11):4120–4124. doi: 10.1073/pnas.87.11.4120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mueh J. R., Herbst K. D., Rapaport S. I. Thrombosis in patients with the lupus anticoagulant. Ann Intern Med. 1980 Feb;92(2 Pt 1):156–159. doi: 10.7326/0003-4819-92-2-156. [DOI] [PubMed] [Google Scholar]
  25. Nilsson I. M., Astedt B., Hedner U., Berezin D. Intrauterine death and circulating anticoagulant ("antithromboplastin"). Acta Med Scand. 1975 Mar;197(3):153–159. doi: 10.1111/j.0954-6820.1975.tb04897.x. [DOI] [PubMed] [Google Scholar]
  26. Nimpf J., Bevers E. M., Bomans P. H., Till U., Wurm H., Kostner G. M., Zwaal R. F. Prothrombinase activity of human platelets is inhibited by beta 2-glycoprotein-I. Biochim Biophys Acta. 1986 Oct 29;884(1):142–149. doi: 10.1016/0304-4165(86)90237-0. [DOI] [PubMed] [Google Scholar]
  27. Oosting J. D., Derksen R. H., Hackeng T. M., van Vliet M., Preissner K. T., Bouma B. N., de Groot P. G. In vitro studies of antiphospholipid antibodies and its cofactor, beta 2-glycoprotein I, show negligible effects on endothelial cell mediated protein C activation. Thromb Haemost. 1991 Dec 2;66(6):666–671. [PubMed] [Google Scholar]
  28. Schousboe I. Characterization of the interaction between beta 2-glycoprotein I and mitochondria, platelets, liposomes and bile acids. Int J Biochem. 1983;15(12):1393–1401. doi: 10.1016/0020-711x(83)90070-8. [DOI] [PubMed] [Google Scholar]
  29. Schousboe I. In vitro activation of the contact activation system (Hageman factor system) in plasma by acidic phospholipids and the inhibitory effect of beta 2-glycoprotein I on this activation. Int J Biochem. 1988;20(3):309–315. doi: 10.1016/0020-711x(88)90356-4. [DOI] [PubMed] [Google Scholar]
  30. Schousboe I., Rasmussen M. S. The effect of beta 2-glycoprotein I on the dextran sulfate and sulfatide activation of the contact system (Hageman factor system) in the blood coagulation. Int J Biochem. 1988;20(8):787–792. doi: 10.1016/0020-711x(88)90065-1. [DOI] [PubMed] [Google Scholar]
  31. Schousboe I. beta 2-Glycoprotein I: a plasma inhibitor of the contact activation of the intrinsic blood coagulation pathway. Blood. 1985 Nov;66(5):1086–1091. [PubMed] [Google Scholar]
  32. Stenflo J. Structure and function of protein C. Semin Thromb Hemost. 1984 Apr;10(2):109–121. doi: 10.1055/s-2007-1004413. [DOI] [PubMed] [Google Scholar]
  33. Wurm H. beta 2-Glycoprotein-I (apolipoprotein H) interactions with phospholipid vesicles. Int J Biochem. 1984;16(5):511–515. doi: 10.1016/0020-711x(84)90168-x. [DOI] [PubMed] [Google Scholar]
  34. van Hinsbergh V. W., Bertina R. M., van Wijngaarden A., van Tilburg N. H., Emeis J. J., Haverkate F. Activated protein C decreases plasminogen activator-inhibitor activity in endothelial cell-conditioned medium. Blood. 1985 Feb;65(2):444–451. [PubMed] [Google Scholar]
  35. van der Meer F. J., van Tilburg N. H., van Wijngaarden A., van der Linden I. K., Briët E., Bertina R. M. A second plasma inhibitor of activated protein C: alpha 1-antitrypsin. Thromb Haemost. 1989 Sep 29;62(2):756–762. [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES