Tissue Factor Activity of Syncytiotrophoblast Plasma Membranes and Tumoral Trophoblast Cells in Culture

Summary

During pregnancy, important modifications of hemostasis occur resulting in mothers in hypercoagulability and the role of placental cells such as trophoblast cells has been hypothesized. In this study, we first showed that syncytiotrophoblast plasma membranes, isolated from normal human placenta, expressed a strong tissue factor (TF) activity. We then studied TF activity of two continuous trophoblast cell lines (JEG-3 and BeWo) in comparison to human umbilical vein endothelial cells (HUVEC) and transformed human endothelial cells (ECV-304). TF assays were performed on intact detached confluent cells. Unstimulated JEG-3 and BeWo cells exhibited a very high TF activity which slightly increased after 2 to 4 h TNF-α stimulation. In contrast, HUVEC and ECV-304 had a lower basal TF activity which was mainly inducible by TNF-a, with a maximum effect after 4 to 6 h stimulation. For both cell types, TF activity was decreased to basal value after 16-hour TNF-α stimulation. These results support that trophoblast cells are able to express TF but the involvement of this property in the hemostatic physiological changes observed during pregnancy, remains to be demonstrated.

• References

• 1 Nemerson Y. Tissue factor and hemostasis. J Am Soc Hematol 1988; 71: 1-8
• 2 Edington TS, Mackman N, Brand K, Ruf W. The structural biology of expression and function of tissue factor. Thromb Haemost 1991; 66: 67-79
• 3 Rapaport SI, Rao LV M. Initiation and regulation of tissue factor-dependent blood coagulation. Arterosclerosis and thrombosis 1992; 12: 1111-1121
• 4 Chesterman CN. Vascular endothelium, haemostasis and thrombosis. Blood Reviews 1988; 2: 88-94
• 5 Mantovani A, Bussolino F, Dejana E. Cytokine regulation of endothelial cell function. FASEB J 1992; 6: 2591-2599
• 6 Bevilacqua MP, Pober JS, Majeau GR, Cotran RS, Gimbrone MA. Interleukin 1 (IL-1) induced biosynthesis and cell surface expression of procoagulant activity human vascular endothelial cells. J Exp Med 1984; 160: 618-623
• 7 Bevilacqua MP, Pober JS, Majeau GR, Fiers W, Cotran RS, Gimbrone MA. Recombinant tumor necrosis factor induces procoagulant activity in cultured human vascular endothelium: characterization and comparison with the actions of interleukin 1. Proc Natl Acad Sci USA 1986; 83: 4533-4537
• 8 Lyberg T, Gaidai KS, Evensen SA, Prydz H. Cellular cooperation in endothelial cell thromboplstin synthesis. Br J Haematol 1983; 53: 85-95
• 9 Gaidai KS, Lyberg T, Evensen SA, Nilsen E, Prydz H. Thrombin induces Thromboplastin synthesis in cultured vascular endothelial cells. Thromb Haemost 1985; 54: 373-376
• 10 Chargaff E. The isolation of preparations of thromboplastic protein from human organs. J Biol Chem 1945; 161: 389-394
• 11 Omsjo JH, Oian P, Maltau JM, Osterud B. Thromboplastin activity in amniotic fluid. Gynecol Obstet Invest 1985; 19: 1-5
• 12 Dalaker K, Prydz H. Regulation of thromboplastin synthesis in mouse placental cells in vitro. Thromb Haemost 1985; 50: 831-834
• 13 Dalaker K, Haug E, Prydz H. Mouse trophoblast cells are constitutive producers of thromboplastin (Factor III) in vitro. Thromb Haemost 1985; 54: 438-441
• 14 Drake TA, Morissey JH, Edington TS. Selective cellular expression of tissue factor in human tissues. Am J Pathol 1989; 134: 1087-1097
• 15 Fleck RA, Rao LV M, Rapaport SI, Varki N. Localization of human tissue factor antigen by immunostaining with monospecific, polyclonal antihuman tissue factor antibody. Thromb Res 1990; 59: 421-37
• 16 Faulk WP, Labarrere A, Carson SD. Tissue factor: identification and characterization of cell types in human placentae. Blood 1990; 76: 86-96
• 17 Khalfoun B, Degenne D, Arbeile-Brassart B, Gutman N, Bardos P. Isolation and characterization of syncytiotrophoblast plasma membrane from human placenta. FEBS Letters 1985; 181: 33-38
• 18 Khalfoun B, Lacord-Bonneau M, Degenne D, Clément L, Bardos P. Characterization of the human syncytiotrophoblast plasma membrane associated components. Int J Biochem 1986; 18: 351-360
• 19 Khalfoun B, Degenne D, Crouzat-Reynes G, Bardos P. Effect of human syncytiotrophoblast plasma membrane-soluble extracts on in vitro mitogen- induced lymphocyte proliferation.A possible inhibition mechanism involving the transferrin receptor. J Immunol 1986; 137: 1187-1193
• 20 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951; 192: 265-275
• 21 Faulk WP, Sanderson AL, Temple A. Distribution of MHC antigens in human placenta chorionic villi. Transplant Proc 1977; 9: 1379-1384
• 22 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
• 23 Patillo RA, Gey GO. The establishment of a cell line of human hormone- synthesizing trophoblastic cells in vitro. Cancer Research 1968; 28: 1231-1236
• 24 Kohler PO, Bridson WE. Isolation of hormone-producing clonal lines of human choriocarcinoma. J Clin Endocr 1971; 32: 683-687
• 25 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria J Clin Invest 1973; 52: 2745-2756
• 26 Takahasi K, Sawasaki Y. Rare spontaneously transformed human endothelial cell line provides useful research tool. In Vitro Cell Dev Biol 1992; 28 A 380-382
• 27 Yoshiba Y. Morphological study of the extravillous trophoblast: proliferation and differentiation:. In: Placenta: Basic Research for Clinical Application. Int Conf on Placenta Soma H. ed Karger; Basel, Tokyo: 1991: 166-175
• 28 Yeh IT. Functional and morphologic expressions of trophoblast. Lab Invest 1989; 61: 1-3
• 29 Graham CH, Lala PK. Mechanisms of placental invasion of the uterus and their control. Biochem Cell Biol 1992; 70: 867-874
• 30 Almus FE, Rao LV M, Rapaport SI. Functional properties of factor Vila/ tissue factor formed with purified tissue factor and with tissue factor expressed on cultures endothelial cells. Thromb Haemost 1989; 62: 1067-1073
• 31 Carson SD, Ramsey CA. Tissue factor (coagulation factor III) is present in placental microvilli and cofractionates with microvilli membrane proteins. Placenta 1985; 6: 5-12
• 32 Rao LV M. Tissue factor as a tumor procoagulant. Cancer and Metastasis Reviews 1992; 11: 249-266
• 33 Stirling Y, Woolf L, North WR S, Seghatchian MJ, Meade TW. Haemostasis in normal pregnancy. Thromb Haemost 1984; 52: 176-82