Platelets Stimulate Thromboplastin Synthesis in Human Endothelial Cells

Summary

Human umbilical vein endothelial cells in culture synthesize thromboplastin upon stimulation with phytohaemagglutinin (PHA) or the tumor promotor 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The thromboplastin activity is further strongly enhanced in a time dependent reaction by the presence of gel-filtered platelets or platelet aggregates. This effect was demonstrable at platelet concentrations lower than those normally found in plasma, it may thus be of pathophysiological relevance. The thromboplastin activity increased with increasing number of platelets added. Cycloheximide inhibited the increase, suggesting that de novo synthesis of the protein component of thromboplastin, apoprotein III, is necessary.

When care was taken to remove monocytes no thromboplastin activity and no apoprotein HI antigen could be demonstrated in suspensions of gel-filtered platelets, platelets aggregated with thrombin or homogenized platelets when studied with a coagulation assay and an antibody neutralization technique.

Keywords

Endothelial cells - Thromboplastin synthesis - Blood platelets - Phytohaemagglutinin - Phorbol ester

• References

• 1 Björkerud S, Bondjers G. Arterial repair and atherosclerosis after mechanical injury. I. Permeability and light microscopic characteristics of endothelium in non-atherosclerotic and atherosclerotic lesions. Atherosclerosis 1971; 13: 355-363
• 2 Moore S. Thromboatherosclerosis in normolipemic rabbits. A result of continued endothelial damage. Lab Invest 1973; 29: 478-487
• 3 Harker LA, Ross R, Slichter S, Scott C. Homocysteine-induced atherosclerosis: The role of endothelial cell injury and platelet response in its genesis. J Clin Invest 1976; 58: 731-741
• 4 Ross R, Harker L. Hyperlipidemia and atherosclerosis. Science 1976; 193: 1094-1100
• 5 Gaynor E, Bouvier C, Spaet TH. Vascular lesions; possible pathogenetic basis of the generalized Shwartzman reaction. Science 1970; 170: 986-988
• 6 Minick CR, Murphy GE. Experimental induction of athero-arteriosclerosis by the synergy of allergic injury to arteries and lipid-rich diet. II. Effect of repeatedly injected foreign protein in rabbits fed a lipid-rich cholesterol-poor diet. Am J Pathol 1973; 73: 265-300
• 7 Ross R, Glomset J. The pathogenesis of atherosclerosis. N Engl J Med 1976; 295: 420-425
• 8 Maynard JR, Dreyer BE, Pitlick FA, Nemerson Y. Tissue factor activity of endothelial and other cultured human cells. Blood 1975; 46: 1046
• 9 Maynard JR, Dreyer BE, Stemerman MB, Pitlick FA. Tissue factor coagulant activity of cultured human endothelial and smooth muscle cells and fibroblasts. Blood 1977; 50: 387-396
• 10 Lyberg T, Gaidai KS, Prydz H. Synthesis of thromboplastin protein in endothelial cells. Proc Second Int Symposium on the Biology of the Vascular Endothelial Cell. p. 107 Cambridge: 1981
• 11 Lyberg T, Gaidai KS, Evensen SA, Prydz H. Cellular cooperation in endothelial cell thromboplastin synthesis. Br J Haematol 1983; 53: 85-95
• 12 Edwards RL, Rickies FR. The role of human T cells (and T cell products) for monocytes tissue factor generation. J Immunol 1980; 125: 606-609
• 13 Maynard JR, Smariga PE. Isolation of a protein from platelets which stimulates an increase in both fibrinolytic inhibitory and tissue factor activities of human foreskin fibroblasts. Fed Proc. 1982. 41. 655
• 14 Korn JH, Rickies FR, Ewan VA, Edwards RL, Myers TJ, Onder O. Mononuclear cell modulation of fibroblast procoagulant activity. J Lab Clin Med 1982; 5: 657-668
• 15 Lyberg T, Nilsson K, Prydz H. Synthesis of thromboplastin by U-937 cells. Br J Haematol 1983; 51: 631-641
• 16 Biggs R, Denson KW E, Riesenberg D, McIntyre C. The coagulant activity of platelets. Br J Haematol 1968; 15: 283-296
• 17 King JB. The function of platelet surface in coagulation. Some preliminary observations with two new tests of platelet function. Thrombos Diathes haemorrh 1974; 32: 492-501
• 18 Semeraro N, Vermylen J. Evidence that washed human platelets possess factor X activator activity. Br JHaematoI 1977; 16: 107-115
• 19 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. J Clin Invest 1973; 52: 2745-2756
• 20 Henriksen T, Evensen SA, Føiling Elgjo R, Vefling A. Human fetal endothelial cells in culture. Scand J Haematol 1975; 14: 233-241
• 21 Tangen O, Berman HJ. Gel-filtration of blood platelets: A methodological report. In: Platelet function and thrombosis. Man-nucci PM, Garini S. (eds.) p. 235 New York: Plenum; 1972
• 22 Hvatum M, Prydz H. Studies on tissue thromboplastin. Solubilization with sodium deoxycholate. Biochim Biophys Acta 1966; 130: 92-101
• 23 Hjort PF. Intermediate reactions in the coagulation of blood with tissue thromboplastin. Scand J Clin Invest. 1957. 09. suppl 027
• 24 Markwell MAK, Haas SM, Bieber LL, Tolbert NE. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 1978; 87: 206-210
• 25 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265-275
• 26 Bjørklid E, Storm E. Purification and some properties of the protein component of tissue thromboplastin from human brain. Biochem J 1977; 165: 89-96
• 27 Prydz H, Gladhaug Å, Factor X. Immunological studies. Thrombos Diathes Haemorrh 1971; 25: 157-165
• 28 Niemetz J, Marcus AJ. The stimulatory effect of platelets and platelet membranes on the procoagulant activity of leukocytes. J Clin Invest 1975; 54: 1437-1443
• 29 Rivers RPA, Hathaway WE, Weston WL. The endotoxin-induced coagulant activity of human monocytes. Br J Haematol 1975; 30: 311-316