Increased Levels of Tissue Factor mRNA in Mononuclear Blood Cells of Patients with Primary Antiphospholipid Syndrome

Summary

Antiphospholipid antibodies (aPL) may stimulate tissue factor (TF) expression in cultured endothelial cells and monocytes, but there are discrepancies as to the expression of TF in the patients with antiphospholipid syndrome (APS). By using reverse transcription and polymerase chain reaction amplification, we have analysed TF mRNA accumulation in freshly isolated mononuclear blood cells (MBC) of 14 patients with primary APS (PAPS) and six normal controls. TF mRNA accumulation was low or absent in uncultured MBC from all normal controls, but was elevated in uncultured MBC from nine of the patients as well as in normal MBC incubated with 100 ng/ml lipopolysaccharide (LPS). Mean levels of TF mRNA, as measured by densitometry, were higher in MBC from patients (N = 14) than in those from controls (N = 6, P = 0.009), and in MBC from patients with a history of thrombosis (N = 9) than in those from patients without thrombosis (N = 5, P = 0.02). Uncultured MBC of patients with thrombosis accumulated TF mRNA at similar levels to LPS-treated normal MBC. Increased levels of TF mRNA were found in eight of ten patients with conventional aPL (ie, anti-cardiolipin antibodies [aCL] and/or lupus anticoagulant [LA]) and littleif any accumulation of TF mRNA was observed in three of four patients without aPL at the time of study. These data strongly suggest that circulating monocytes of many patients with PAPS are subjected to an up-regulated TF expression that may well explain their prothrombotic state. Although the presence or absence of TF mRNA in MBC was associated with, respectively, the presence or absence of conventional aPL in 11 of the 14 patients studied, our study cannot exclude the involvement of factors other than aCL or LA in inducing TF expression.

Key Words

Antiphospholipid syndrome - in vivo - monocyte - mRNA - tissue factor

• References

• 1 Roubey RAS, Hoffman M. From antiphospholipid syndrome to antibody-mediated thrombosis. Lancet 1997; 350: 1491-3.
• 2 Blank M, Cohen J, Toder V, Shoenfeld Y. Induction of antiphospholipid syndrome in naive mice with mouse lupus monoclonal and human polyclonal anticardiolipin antibodies. Proc Natl Acad Sci USA 1991; 88: 3069-73.
• 3 Bakimer R, Fishman P, Blank M, Sredni B, Djaldetti M, Shoenfeld Y. Induction of primary antiphospholipid syndrome in mice by immunization with a human monoclonal anticardiolipin antibody (H-3). J Clin Invest 1992; 89: 1558-63.
• 4 Krause I, Blank M, Shoenfeld Y. Immunomodulation of experimental APS: lessons from murine models. Lupus 1996; 5: 458-62.
• 5 Roubey RAS. Immunology of the antiphospholipid antibody syndrome. Arthritis Rheum 1996; 39: 1444-54.
• 6 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 Haemost 1989; 62: 654-60.
• 7 Brandt JT. The effect of lupus anticoagulants on expression of tissue factor activity by cultured endothelial cells. Thromb Haemost 1991; 65: 673 (abstract).
• 8 Oosting JD, Derksen RHWM, Blokzijl L, Sixma JJ, de Groot PG. Antiphospholipid antibody positive sera enhance endothelial cell procoagulant activity: studies in a thrombosis model. Thromb Haemost 1992; 68: 278-84.
• 9 Amengual O, Atsumi T, Khamashta MA, Hughes GRV. The role of the tissue factor pathway in the hypercoagulable state in patients with the antiphospholipid syndrome. Thromb Haemost 1998; 79: 276-81.
• 10 Schved JF, Gris JC, Ollivier V, Wautier JL, Tobelem G, Caen J. Procoagulant activity of endotoxin or tumor necrosis factor activated monocytes is enhanced by IgG from patients with lupus anticoagulant. Am J Hematol 1992; 41: 92-6.
• 11 Kornberg A, Blank M, Kaufman S, Shoenfeld Y. Induction of tissue factor-like activity in monocytes by anti-cardiolipin antibodies. J Immunol 1994; 153: 1328-32.
• 12 Martini F, Farsi A, Gori AM, Boddi M, Fedi S, Domeneghetti MP, Passaleva A, Prisco D, Abbate R. Antiphospholipid antibodies (aPL) increase the potential monocyte procoagulant activity in patients with systemic lupus erythematosus. Lupus 1996; 5: 206-11.
• 13 Reverter J-C, Tàssies D, Font J, Monteagudo J, Escolar G, Ingelmo M, Ordinas A. Hypercoagulable state in patients with antiphospholipid syndrome is related to high induced tissue factor expression on monocytes and to low free protein S. Arterioscler Thromb Vasc Biol 1996; 16: 1319-26.
• 14 Reverter J-C, Tàssies D, Font J, Khamashta MA, Ichikawa K, Cervera R, Escolar G, Hughes GRV, Ingelmo M, Ordinas A. Effects of human monoclonal anticardiolipin antibodies on platelet function and on tissue factor expression on monocytes. Arthritis Rheum 1998; 41: 1420-7.
• 15 Østerud B, Flægstad T. Increased tissue thromboplastin activity in monocytes of patients with menigococcal infection: related to an unfauvorable prognosis. Thromb Haemost 1983; 49: 5-7.
• 16 Mackman N, Morrissey JH, Fowler B, Edgington TS. Complete sequence of the human tissue factor gene, a highly regulated cellular receptor that initiates the coagulation protease cascade. Biochemistry 1989; 28: 1755-62.
• 17 Edwards RL, Rickles FR, Cronlund M. Abnormalities of blood coagulation in patients with cancer: mononuclear cell tissue factor generation. J Lab Clin Med 1981; 98: 917-28.
• 18 Weis JR, Pitas RE, Wilson BD, Rodgers GM. Oxidized low-density lipoprotein increases cultured human endothelial cell tissue factor activity and reduces protein C activation. FASEB J 1991; 5: 2459-65.
• 19 Cuadrado MJ, López-Pedrera Ch, Khamashta MA, Camps MT, Tinahones F, Torres A, Hughes GRV, Velasco F. Thrombosis in primary antiphospholipid syndrome: a pivotal role for monocyte tissue factor expression. Arthritis Rheum 1997; 40: 834-41.
• 20 Albrecht S, Kotzsch M, Siegert G, Luther T, Großmann H, Großer M, Müller M. Detection of circulating tissue factor and factor VII in a normal population. Thromb Haemost 1996; 75: 772-7.
• 21 Williams FMK, Jurd K, Hughes GRV, Hunt BJ. Antiphospholipid syndrome patients’ monocytes are “primed” to express tissue factor. Thromb Haemost 1998; 80: 864-5.
• 22 Van Ginkel CJW, Van Aken WG, Oh JIH, Vreeken J. Stimulation of monocyte procoagulant activity by adherence to different surfaces. Br J Haematol 1977; 37: 35-45.
• 23 Harris EN, Baguley E, Asherson RA, Hughes GRV. Clinical and serological features of the antiphospholipid syndrome (APS). Br J Rheumatol 1987; 26: 19 (abstract).
• 24 Brandt JT, Triplett DA, Alving B, Scharrer I. Criteria for the diagnosis of lupus anticoagulants: an update. Thromb Haemost 1995; 74: 1185-90.
• 25 Verhagen HJM, Heijnen-Snyder GJ, Vink T, Pronk A, van Vroonhoven TJMV, Eikelboom BC, Sixma JJ, de Groot PG. Tissue factor expression on mesothelial cells is induced during in vitro culture: manipulation of culture conditions creates perspectives for mesothelial cells as a source for cell seeding procedures on vascular grafts. Thromb Haemost 1995; 74: 1096-102.
• 26 de Prost D, Ollivier V, Ternisien C, Chollet-Martin S. Increased monocyte procoagulant activity independent of the lupus anticoagulant in patients with systemic lupus erythematosus. Thromb Haemost 1990; 64: 216-21.
• 27 Haskill S, Johnson C, Eierman D, Becker S, Warren K. Adherence induces selective mRNA expression of monocyte mediators and proto-oncogenes. J Immunol 1988; 140: 1690-4.
• 28 Fuhlbrigge RC, Chaplin DD, Kiely J-M, Unanue ER. Regulation of interleukin 1 gene expression by adherence and lipopolysaccharide. J Immunol 1987; 138: 3799-802.
• 29 Greaves M, Hill MB, Phipps J, Hughes P, Ford I. The pathogenesis of the antiphospholipid syndrome. Thromb Haemost 1996; 76: 817-8.
• 30 Vermylen J, Hoylaerts MF, Arnout J. Antibody-mediated thrombosis. Thromb Haemost 1997; 78: 420-6.