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Thromb Haemost 1992; 68(03): 278-284
DOI: 10.1055/s-0038-1656365
Original Article
Schattauer GmbH Stuttgart

Antiphospholipid Antibody Positive Sera Enhance Endothelial Cell Procoagulant Activity – Studies in a Thrombosis Model

Janine D Oosting
1   The Department of Haematology, University Hospital Utrecht, The Netherlands
2   The Department of Internal Medicine (Division of Clinical Immunology and Infectious diseases), University Hospital Utrecht, The Netherlands
,
Ronald H W M Derksen
2   The Department of Internal Medicine (Division of Clinical Immunology and Infectious diseases), University Hospital Utrecht, The Netherlands
,
Laya Blokzijl
1   The Department of Haematology, University Hospital Utrecht, The Netherlands
2   The Department of Internal Medicine (Division of Clinical Immunology and Infectious diseases), University Hospital Utrecht, The Netherlands
,
Jan J Sixma
1   The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Philip G de Groot
1   The Department of Haematology, University Hospital Utrecht, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 16 January 1992

Accepted after revision 01 April 1992

Publication Date:
04 July 2018 (online)

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Summary

The effect of sera and IgG from 12 patients with systemic lupus erythematosus (SLE) on the endothelial cell (EC) procoagulant activity (PCA) was investigated in an in vitro thrombosis model. Six of the 12 SLE sera contained antiphospholipid antibodies (aPL).

EC were stimulated for 8 h at 37° C with or without 50 pM tumor necrosis factor (TNF) in culture medium containing 20% patient or control serum. Then the endothelial cell matrix (ECM) was isolated and subsequently exposed in a perfusion chamber to circulating normal whole blood, anticoagulated with low molecular weight heparin (LMWH). The PCA of the ECM was determined as the amount of generated fibrinopeptide A (FPA) in samples taken before and after perfusion. Furthermore, cross sections were made of the perfused matrix and analyzed for platelet adhesion and aggregate formation.

All six aPL containing sera induced a small, but significant increase of ECM procoagulant activity. When added in combination with a low dose of TNF (50 pM), a synergistic enhancement of ECM procoagulant activity was found. The FPA generation was increased to 150–614% from the values obtained after stimulation with TNF and control serum. Also a shift towards the formation of larger platelet thrombi was observed. After stimulation with TNF and patient serum the surface of ECM covered with large aggregates (> 5 µm) was increased by 124–329% compared to the results obtained after stimulation with control serum and TNF. When patient sera were depleted from IgG the effects were strongly decreased.

These data show that the potentiation of TNF-induced PCA formation by aPL containing sera from patients with SLE leads to enhanced thrombus formation in an in vitro thrombosis model. This may help explain the increased thrombotic tendency in these patients.

 

  • References

  • 1 Harris EN, Gharavi AE, Boey ML, Patel BM, Mackworth-Young CG, Loizou S, Hughes GRV. Anticardiolipin antibodies: detection by radioimmunoassay and association with thrombosis in systemic lupus erythematosus. Lancet 1983; 2: 1211-1214
    PubMedGoogle Scholar
  • 2 Derksen RHWM, Kater L. Lupus anticoagulant: revival of an old phenomenon. Clin Exp Rheumatol 1985; 3: 349-357
    PubMedGoogle Scholar
  • 3 Feinstein DI. Lupus anticoagulant, thrombosis and fetal loss. N Engl J Med 1985; 313: 1348-1350
    CrossrefPubMedGoogle Scholar
  • 4 Derksen RHWM, Bouma BN, Kater L. The striking association between lupus anticoagulant and fetal loss in systemic lupus erythematosus. Arthritis Rheum 1986; 29: 695-696
    PubMedGoogle Scholar
  • 5 Lechner K, Pabinger-Fasching I. Lupus anticoagulants and thrombosis. Haemostasis 1985; 15: 254-262
    PubMedGoogle Scholar
  • 6 Comp PC, De Bault LE, Esmon NE, Esmon CP. Human thrombomodulin is inhibited by IgG from two patients with non-specific anticoagulants. Blood 1983; 62: 1099
    PubMedGoogle 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 Haemostas 1988; 60 (1) 54-58
    PubMedGoogle Scholar
  • 8 Hasselaar P, Derksen RHWM, Blokzijl L, De Groot PG. Thrombosis associated with antiphospholipid antibodies cannot be explained by effects on endothelial and platelet prostanoid synthesis. Thromb Haemostas 1988; 59: 80-85
    PubMedGoogle Scholar
  • 9 Francis RB, McGehee WG, Feinstein DI. Endothelial-dependent fibrinolysis in subjects with the lupus anticoagulant and thrombosis. Thromb Haemostas 1988; 59: 412-414
    PubMedGoogle Scholar
  • 10 Derksen RHWM, Hasselaar P, Oosting JD, De Groot PG. The antiphospholipid antibody dilemma. Clin Rheumatol 1990; 9 No. 1, Suppl 1 39-44
    CrossrefPubMedGoogle Scholar
  • 11 Oosting JD, Derksen RHWM, Hackeng TM, Van Vliet M, Preissner KT, Bouma BN, De Groot PG. In vitro studies of antiphospholipid antibodies and its cofactor, β2-glycoprotein I, show negligible effects on endothelial cell mediated protein C activation. Thromb Haemostas 1991; 66: 666-671
    PubMedGoogle Scholar
  • 12 Hasselaar P, Derksen RHWM, Oosting JD, Blokzijl L, De Groot PG. Synergistic effect of low doses of tumor necrosis factor and sera from patients with systemic lupus erythematosus on the expression of procoagulant activity by cultured endothelial cells. Thromb Haemostas 1989; 62: 654-660
    PubMedGoogle Scholar
  • 13 Zwaginga JJ, Sixma JJ, De Groot PG. Activation of endothelial cells with various stimuli induces platelet thrombus formation on their matrix: studies of a new in vitro thrombosis model using low molecular weight heparin anticoagulated whole blood. Arteriosclerosis 1990; 10: 49-61
    CrossrefPubMedGoogle Scholar
  • 14 Tan EM, Cohen AS, Fries JF. The 1982 revised criteria for the classification of SLE. Arthritis Rheum 1982; 25: 1271-1277
    CrossrefPubMedGoogle Scholar
  • 15 Derksen RHWM, Hasselaar P, Blokzijl L, Gmelig Meyling FHJ, De Groot PG. Coagulation screen is more specific than the anticardiolipin antibody ELISA in defining a thrombotic subset of lupus patients. Ann Rheum Dis 1988; 47: 364-371
    CrossrefPubMedGoogle Scholar
  • 16 Rosner E, Pauzner R, Lusky A, Modan M, Many A. Detection and quantitative evaluation of lupus circulating anticoagulant activity. Thromb Haemostas 1987; 57: 144-147
    PubMedGoogle Scholar
  • 17 Harris EN. The second international anti-cardiolipin standardization workshop/The Kingston anti-phospholipid antibody study (KAPS) group. Am J Clin Pathol 1990; 94: 476-484
    CrossrefPubMedGoogle Scholar
  • 18 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. J Clin Invest 1973; 52: 2745-2756
    CrossrefPubMedGoogle Scholar
  • 19 Willems C, Astaldi GCB, De Groot PG, Janssen MC, Gonsalves MD, Zeijlemaker WP, Van Mourik JA, Van Aken WG. Media conditioned by cultured human vascular endothelial cells inhibit the growth of vascular smooth muscle cells. Exp Cell Res 1982; 139: 191-197
    CrossrefPubMedGoogle Scholar
  • 20 Ryan J, Brett J, Kisiel W, Stern D. Tumor necrosis factor (TNF)-induced endothelial cell tissue factor activity is predominantly associated with the subendothelial matrix. Thromb Haemostas 1991; 65: 702
    PubMedGoogle Scholar
  • 21 Houdijk WPM, De Groot PG, Nievelstein PFEM, Sakariassen KS, Sixma JJ. Von Willebrand factor and fibronectin but not thrombospondin are involved in platelet adhesion to the extracellular matrix of human vascular endothelial cells. Arteriosclerosis 1986; 6: 24-33
    CrossrefPubMedGoogle Scholar
  • 22 Sakariassen KS, Bolhuis PA, Sixma JJ. Platelet adherence to subendothelium of human arteries in pulsatile and steady flow. Thromb Res 1980; 19: 547-559
    CrossrefPubMedGoogle Scholar
  • 23 Sakariassen KS, Aarts PAMM, De Groot PG, Houdijk WPM, Sixma JJ. A perfusion chamber developed to investigate platelet interaction in flowing blood with human vessel wall cells, their extracellular matrix and purified components. J Lab Clin Med 1983; 102: 522-535
    PubMedGoogle Scholar
  • 24 Sakariassen KS, Bolhuis PA, Sixma JJ. Human blood platelet adhesion to artery subendothelium is mediated by factor VIII-von Willebrand factor bound to the subendothelium. Nature 1979; 279: 636-638
    CrossrefPubMedGoogle Scholar
  • 25 Baumgartner HR. The role of blood flow in platelet adhesion, fibrin deposition, and formation of mural thrombi. Microvasc Res 1973; 5: 167-179
    CrossrefPubMedGoogle Scholar
  • 26 Nemerson Y. Tissue factor and haemostasis. Blood 1988; 71: 01-12
    PubMedGoogle Scholar
  • 27 Tijburg PNM, Ryan J, Stern DM, Wollitzky B, Rimon S, Rimon B, Handley D, Nawroth PP, Sixma JJ, De Groot PG. Activation of the coagulation mechanism on TNF-stimulated cultured endothelial cells and their extracellular matrix. Role of flow and factor IX/IXa. J Biol Chem 1991; 266: 12067-12074
    PubMedGoogle Scholar
  • 28 Bach R, Rifkin DB. Expression of tissue factor procoagulant activity: Regulation by cytosolic calcium. Proc Natl Acad Sci USA 1990; 87: 6995-6999
    CrossrefPubMedGoogle Scholar
  • 29 Hasselaar P, Derksen RHWM, Blokzijl L, De Groot PG. Crossreactivity of antibodies directed against cardiolipin, DNA, endothelial cells and blood platelets. Thromb Haemostas 1990; 63: 169-173
    PubMedGoogle Scholar
  • 30 Maury CPJ, Teppo A-M. Tumor necrosis factor in the serum of patients with systemic lupus erythematosus. Arthritis Rheum 1989; 32: 146-150
    CrossrefPubMedGoogle Scholar
  • 31 Maury CP, Teppo A-M. Cachectin/tumor necrosis factor-alpha in the circulation of patients with rheumatic disease. Int J Tissue React 1989; 11: 189-193
    PubMedGoogle Scholar
  • 32 Malave I, Searles RP, Montano J, Williams Jr RC. Production of tumor necrosis factor/cachectin by peripheral blood mononuclear cells in patients with systemic lupus erythematosus. Int Arch Allergy Appl Immunol 1989; 89: 355-361
    CrossrefPubMedGoogle Scholar
  • 33 Quadros NP, Roberts-Thomson PJ, Gallus AS. IgG and IgM antiendothelial cell antibodies in patients with collagen-vascular disorders. Rheumatol Int 1990; 10: 113-119
    CrossrefPubMedGoogle Scholar
  • 34 Malia RG, Kitchen S, Greaves M, Preston FE. Inhibition of activated protein C and its cofactor protein S by antiphospholipid antibodies. Br J Haematol 1990; 76: 101-107
    CrossrefPubMedGoogle Scholar
  • 35 Marciniak E, Romond E. Impaired catalytic function of activated protein C: a new in vitro manifestation of lupus anticoagulant. Blood 1989; 74: 2426-2432
    PubMedGoogle Scholar
  • 36 Tannenbaum SH, Finko R, Cines DB. Antibody and immune complexes induce tissue factor production by human endothelial cells. J Immunol 1986; 137: 1532-1537
    PubMedGoogle Scholar
  • 37 Rustin MHA, Bull HA, Machin SJ, Isenberg DA, Snaith ML, Dowd PM. Effects of the lupus anticoagulant in patients with systemic lupus erythematosus on endothelial cell prostacyclin release and procoagulant activity. J Invest Dermatol 1988; 90: 744-748
    CrossrefPubMedGoogle Scholar