Effect of Lupus Anticoagulant on Antithrombogenic Properties of Endothelial Cells - Inhibition of Thrombomodulin-Dependent Protein C Activation

Summary

We have investigated the effect of purified immunoglobulin G (IgG) on endothelial cell functions in 16 patients with lupus anticoagulant, 9 of whom had systemic lupus erythematosus (SLE). Spontaneous or thrombin-stimulated secretion of prostacyclin (PGI₂) by cultured human endothelial cells from umbilical cord vein (HUVEC) was not inhibited by the patient’s IgG. Nor was spontaneous release of tissue plasminogen activator (t-PA) or of its inhibitor (PAI) modified in the presence of patient’s IgG.

The rate of activation of purified protein C (PC) by HUVEC in the presence of thrombin was significantly lowered by patient’s IgG or Fab’ fragment (inhibition of 43%). Neutralization of this effect was obtained by incubation of a greater quantity of phospholipids (phosphatidylcholine, phosphatidylserine) with the patient’s IgG. Activation of PC was also performed using purified rabbit thrombomodulin (TM) and a similar inhibition of the patient IgG was observed (inhibition of 48%) but the activation of Gla-domainless PC was not modified.

Key words

Lupus anticoagulant - Endothelial cells - Protein C - Thrombomodulin - Prostacyclin

• References

• 1 Frich PG. Acquired circulating anticoagulants in systemic “collagen disease” autoimmune thromboplastin deficiency. Blood 1955; 10: 691-693
• 2 Harris EN, Gharave AE, Boey ML, Patel BM, Mac Kunth Young CG, Loizou S, Hughes GRV. Anticardiolipin antibodies: detection by radioimmunoassay and association with thrombosis in systemic lupus erythematosus. Lancet 1983; II: 1211-1214
• 3 Shoenfeld Y, Rauch J, Massicatte H, Data SK, Schwartz J, Stollar D, Schwartz AS. Polyspecificity of monoclonal lupus antibodies produced by human hybridromas. N Engl J Med 1983; 308: 414-420
• 4 Mueh JR, Herbst KD, Rapaport SI. Thrombosis in patients with the lupus anticoagulant. Ann Intern Med 1980; 92: 156-159
• 5 Lechner K. Acquired inhibitors in non hemophilic patients. Haemostasis 1984; 3: 65-93
• 6 Boey ML, Colaco CB, Charair AE, Elkon KB, Loizou S, Hugues GRV. Thrombosis in SLE: striking association with the presence of circulating lupus anticoagulant. Br Med J 1983; 287: 1021-1023
• 7 Carreras LO, Defreyn G, Machin SJ, Vermylen J, Deman R, Spitz B, Van AsscheA. Arterial thrombosis, intrauterine death and “lupus” anticoagulant: detection of immunoglobulin interfering with pros-tacyclin formation. Lancet 1981; I: 244-246
• 8 Cines DB, Lyss AP, Reeber M, Bian M, Deharatus J. Presence of complement fixing antiendothelial cell antibodies in SLE. J Clin Invest 1984; 73: 611-625
• 9 Ryken DC, Van Hinsberg VW M. Quantitation of tissue plasminogen activator in human endothelial cell cultures by use of an enzyme immunoassay. Thromb Res 1984; 33: 145-153
• 10 Owen WG, Esmon CT. Functionnal properties of an endothelial cell cofactor for thrombin catalized activation of protein C. J Biol Chem 1981; 256: 5232-5535
• 11 Esmon NL, De Bault LE, Esmon CT. Proteolytic formation and properties of carboxyglutamic acid domain less protein C. J Biol Chem 1983; 258: 5548-5553
• 12 Esmon NL, Owen WG, Esmon CT. Isolation of a membrane bound cofactor for thrombin catalyzed activation of protein C. J Biol Chem 1982; 257: 859-864
• 13 Caen J, Larrieu MJ, Samama M. L’hémostase: méthodes d’exploration et diagnostic pratique. L’Expansion Scientifique; Paris: 1975. 1 Vol
• 14 Hardisty RM, Ingram GI C. Bleeding Disorders; Blackwell, Oxford: 1965: 315 pp
• 15 Goudswaard J, Van Der Donk JA, Noordzij A, Van Dam RH, Vaerman J. Protein A reactivity of various mammalian immunoglobulins. Scand J Immunol 1978; 8: 21-28
• 16 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria J Clin Invest 1983; 52: 2745-2756
• 17 Pradelles P, Gassi J, Maclouf J. Enzyme immunoassays of eicosanoids using acetylcholine esterase as label: an alternative to radioimmunoassay. Anal Chem 1985; 57: 1170-1173
• 18 Engwall E, Carlsson HE. Enzyme linked immunoadsorbent assay Elisa. In: First International Symposium on Immunoenzymatic Technique INSERM Symposium No. 2. Feldmann M. (ed) North-Holland Publishing Company; Amsterdam: 1976. pp 135-147
• 19 Wiman B, Melbring G, Ranby M. Plasminogen activator release during venous stasis and exercise as determined by a new specific assay. Clin Chim Acta 1982; 127: 279-280
• 20 Verheijen JH, Chang GT G, Kluft C. Evidence for the occurrence of a fast-acting inhibitor for tissue type plasminogen activator in human plasma. Thromb Haemostas 1984; 51: 392-395
• 21 Johnson AJ, Esmon NL, Lane TM, Esmon CT. Structural charges required for activation of protein C are induced by Ca⁺⁺ binding to a high affinity site that does not contain carboxyglutamic acid. J Biol Chem 1983; 258: 5554-5560
• 22 Salem HH, Maruyana I, Ishii H, Majerus PW. Isolation and characterization of thrombomodulin from human placenta. J Biol Chem 1984; 259: 12246-12251
• 23 Shapiro SS, Thiagarajan P. Lupus anticoagulants. Prog Thromb Haemostas 1982; 6: 263-283
• 24 Kelsey PR, Stevenson KJ, Poller L. The diagnosis of lupus anticoagulants by the activated partial thromboplastin time. The central role of phosphotidylserine. Thromb Haemostas 1984; 52: 172-175
• 25 Thiagarjan P, Shapiro SS, De Marco L. Monoclonal immunoglobulin M coagulation inhibitor with phospholipid specificity. J Clin Invest 1980; 66: 397-405
• 26 Goldsmith JC, Cormick JJ. Immunologic injury to vascular endothelial cells. Effect on release of prostacyclin Blood 1984; 63: 984-989
• 27 Fitzgerald GA, Smith B, Perdersen AK, Brash AR. Increased prostacyclin biosynthesis in patients with severe atherosclerosis and platelet activation. N Engl J Med 1984; 310: 1065-1068
• 28 Wautier JL, Pintigny D, Maclouf J, Wautier MP, Corvazier E, Caen J. Realease of prostacyclin after erythrocyte adhesion to cultured vascular endothelium. J Lab Clin Med 1986; 107: 210-215
• 29 Angles Cano E, Sultan Y, Clauvel JP. Predisposing factors to thrombosis in SLE possible relation to endothelial cell damage. J Lab Clin Med 1979; 94: 312-323
• 30 Glas Greenwalt P, Shashi Kant K, Allen C, Pollek VE. Fibrinolysis in health and disease: severe abnormalities in SLE. J Lab Clin Med 1984; 104: 962-976
• 31 Malar RA, Kleiss AJ, Griffin JH. Mechanism of action of human activated protein C, a thrombin dependent anticoagulant enzyme. Blood 1982; 59: 1067-1072
• 32 Van Hinsbergh VW N, Bertessa RM, Van Wijn Gaarden A, Van Tilburg NH, Fineis JJ, Hawerkate F. Activated protein C decreases plasminogen activator-inhibitor activity in endothelial cell conditioned medium. Blood 1985; 65: 444-451
• 33 Salem HH, Esmon NL, Esmon CT, Mayers PW. Effects of thrombomodulin and coagulation factor Va light chain on protein C activation in vitro. J Clin Invest 1984; 73: 968-972
• 34 Nixon RR, Comp PC, Esmon CT. Role of Gla-domain in activated protein C. Fed Proc 1983; 42: 1993 (abstract)
• 35 Griffin JH, Evatt B, Zimmerman TS, Kleiss AJ, Wideman C. Deficiency of protein C in congenital thrombotic disease. J Clin Invest 1980; 68: 1370-1373
• 36 Freyssinet JM, Brami B, Gauchy J, Cazenave JP. Coextraction of thrombomodulin and tissue factor from human placenta. Effects of concanavalin A and phospholipid environment on activity. Thromb Haemostas 1986; 55: 112-118
• 37 Freyssinet JM, Wiesel ML, Gauchy J, Boneu B, Cazenave JP. An IgM lupus anticoagulant that neutralizes the enhancing effect of phospholipid on purified endothelial thrombomodulin activity. A mechanism for thrombosis Thromb Haemostas 1986; 55: 309-313