Expression and Localization of Tissue Factor-Based Procoagulant Activity (PCA) in Pigeon Monocyte-Derived Macrophages

Summary

Monocyte-derived macrophages, focal to initiation and progression of atherosclerosis, have been implicated in thrombotic complication of this disease. In the present study we demonstrated tissue factor based procoagulant activity in cultured macrophages from the White Carneau pigeon following endotoxin (1-2 μg/ml) stimulation. This macrophage procoagulant activity paralleled activity obtained with pigeon brain homogenate. We used Enzyme-Linked Coagulation Assay (ELCA), an ultrasensitive microtiter plate assay, to measure procoagulant activity in these cells. Through the use of clotting factors purified from pigeon plasma, procoagulant activity could be detected with as few as 1-3 cells. Tissue factor antigen, detected through the use of immunogold labelling in conjunction with a polyclonal antibody which was highly specific to human tissue factor, was distributed uniformly over the plasma membrane of the endotoxin-stimulated cells. These studies suggest that this procoagulant activity might play an important role in the pathobiology of atherosclerosis in White Carneau pigeons by initiating fibrin polymerization and thus leading to thrombotic complications of the disease.

• References

• 1 Lewis JC, Taylor RG, Jerome WG. Foam cell characteristics in coronary arteries and aortas of White Cameau pigeons with moderate hypercholesterolemia. Ann NY Acad Sci 1985; 454: 091-100
• 2 Ross R. The pathogenesis of atherosclerosis: an update. N Engl J Med 1986; 326: 488-500
• 3 Davies PF. Vascular cell interactions with special reference to pathogenesis of atherosclerosis. Lab Invest 1986; 55: 05-26
• 4 Nawroth PP, Handley DA, Esmon CT, Stem DM. Interleukin-1 induces endothelial cell procoagulant while suppressing cell surface anticoagulant activity. Proc Natl Acad Sci 1986; 83: 3460-3464
• 5 Bevilacqua MP, Pober JS, Wheeler ME, Costran RS, Gimbrone MA. Interleukin-1 activation of vascular endothelium. Effect on procoagulant activity and leukocyte adhesion. Am J Pathol 1985; 121: 393-403
• 6 Smith EB, Alexander KM, Massie IB. Insoluble fibrin in human aortic intima. Quantitation studies on the relationship between insoluble fibrin, soluble fibrinogen and low density lipoprotein. Atherosclerosis 1976; 23: 19-39
• 7 Smith EB. Fibrinogen, fibrin and fibrin degradation products in relation to atherosclerosis. In: Atherosclerosis VII. Fidge NH, Nestel PJ. eds Amsterdam: Elsevier Science Publishers BV; 1986: 459-462
• 8 Geczy CL. Induction of macrophage procoagulant by products of activated lymphocyte. Haemostasis 1984; 14: 400-411
• 9 Semeraro N, Biondi A, Lorenzet R, Locati D, Mantovani A, Donati MB. Direct induction of tissue factor synthesis by endotoxin in human macrophages from diverse anatomical sites. Immunology 1983; 50: 529-535
• 10 Semeraro N, Montemurro P, Conese M, Giordano D, Stella M, Restaino A, Cagnazzo G, Colucci M. Procoagulant activity of mononuclear phagozytes from different anatomical sites in patients with gynecological malignancies. Int J Cancer 1990; 45: 251-254
• 11 Lyberg T, Prydz H. Phorbol esters induce synthesis of thromboplastin activity in human monocytes. Biochem J 1981; 94: 699-706
• 12 Semeraro N, DeLucia O, Lattanzio A, Montemurro P, Domenico G, Loizzi M, Carpagnano F. Procoagulant activity of human alveolar macrophages: different expression in patients with lung cancer. Int J Cancer 1986; 37: 525-529
• 13 Zeldis SM, Nemerson Y, Pitlick FA, Lentz TL. Tissue factor (thromboplastin): localization to plasma membranes by peroxidase-conjugated antibodies. Science 1972; 175: 766-768
• 14 Tipping PG, Malliaros J, Holdsworth SR. Procoagulant activity expression by macrophages from atheromatous vascular plaques. Atherosclerosis 1989; 79: 237-243
• 15 Wilcox JN, Smith KM, Schwartz SM, Gordon D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci 1989; 86: 2839-2843
• 16 Edwards RL, Rickies FR. Macrophage procoagulants. In: Progress in Haemostasis and Thrombosis Vol. 7. Spact TH. ed. New York: Grune & Stratton; 1984: 183-201
• 17 Bach RR. Initiation of coagulation by tissue factor. CRC Crit Rev Biochem 1988; 23: 339-368
• 18 Nemerson Y, Bach R. Tissue factor revisited. Prog Hemostas Thromb 1982; 6: 237-261
• 19 Osterud B, Rapaport SI. Activation of factor IX by the reaction product of tissue factor and factor VII: additional pathway for initiating blood coagulation. Proc Natl Acad Sci USA 1977; 74: 5260-5264
• 20 Clarkson TB, Prichard RW, Netsky MG, Lofland HB. Atherosclerosis in pigeons. AMA Arch Pathol 1959; 68: 143-147
• 21 Prichard RW, Clarkson TB, Goodman HO, Lofland HB. Aortic atherosclerosis in pigeons and its complications. AMA Arch Pathol 1964; 77: 244-257
• 22 Jones NL, Gupta M, Deckelman C, Hantgan RR, Lewis JC. Monocyte/macrophages: a potential role in thrombosis secondary to atherosclerosis. Arteriosclerosis 1990; 10: A822
• 23 Gupta M, Jones NL, Hantgan RR, Lewis JC. Pigeon monocyte derived macrophages express glycoproteins Ilb/IIIa. Arteriosclerosis 1990; 10: A822
• 24 Jones NL, Allen NS, Lewis JC. βVLDL uptake by pigeon monocyte derived macrophages. Correlation of binding dynamics with 3dimensional ultrastructure. Cell Motil Cytoskel 1991; 19: 139-151
• 25 Jones NL, Lewis JC, Kilpatrick BA. Cytoskeletal disruption during human cytomegalovirus infection of human lung fibroblasts. Eur J Cell Biol 1986; 41: 304-312
• 26 Doellgast GJ, Rothberger H. Enzyme-linked coagulation assay. II. A sensitive assay for tissue factor and factors II, VII, and XI. Anal Biochem 1986; 152: 199-207
• 27 Pitlick FA, Nemerson Y. Purification and characterization of tissue factor apoprotein. Meth Enzymol 1976; 45: 37-48
• 28 Doellgast GJ. Enzyme-linked coagulation assay. IV. Sensitive sandwich enzyme-linked immunosorbent assays using Russell’s viper venom factor X activator-antibody conjugates. Anal Biochem 1987; 167: 097-105
• 29 Miletich JP, Broze Jr GJ, Majerus PW. The synthesis of sulphated dextran beads for isolation of human plasma coagulation factors II, IX and X. Anal Biochem 1980; 105: 304-310
• 30 Miletich JP, Broze Jr GJ, Majerus PW. Purification of human coagulation factors II, IX and X using sulphated dextran beads. In: Methods in Enzymology. Lorand L. ed. New York: Academic Press; 1981: 221-228
• 31 Broze GJ. Majerus PW Purification and properties of human coagulation factor VII. J Biol Chem 1980; 255: 1242-1247
• 32 Hetland O, Brovold AB, Holme R, Gaudernack G, Prydz H. Thromboplastin (tissue factor) in plasma membranes of human monocytes. Biochem J 1985; 228: 735-743
• 33 Surprenant YM, Zuckerman SH. A novel microtiter plate assay for the quantitation of procoagulant activity on adherent monocytes, macrophage and endothelial cells. Thromb Res 1989; 53: 339-346
• 34 Prydz H, Allison AC. Tissue thromboplastin activity of isolated human monocytes. Thromb Haemostas 1978; 39: 582-591
• 35 Schuff-Werner P, Claus G, Armstrong VW, Kostering H, Seidel D. Enhanced procoagulatory activity (PCA) of human monocytes/macrophages after in vitro stimulation with chemically modified LDL. Atherosclerosis 1989; 78: 109-112
• 36 Lyberg T, Gaidal KS, Evenson SA, Prydz H. Cellular cooperation in endothelial cell thromboplastin synthesis. Brit J Haematol 1983; 53: 85-95
• 37 Osterud B, Bjorklid E. The production and availability of tissue thromboplastin in cellular populations of whole blood exposed to various concentrations of endotoxin. An assay for detection of endotoxin. Scand J Haematol 1982; 29: 175-184
• 38 Kaplan JE, Cardarelli PM, Rourke FJ, Weston LK, Moon DG, Blumenstock FA. Fibronectin augments binding of fibrin to macrophages. J Lab Clin Med 1989; 113: 168-176
• 39 Owen J, Grossman BA, Palmer RH. Hyperlipidemia and in vivo hemostatic system activation. Sem Thromb Hemostas 1988; 14: 241-245
• 40 Thompson WD, Smith EB. Atherosclerosis and the coagulation system. J Pathol 1989; 159: 097-106
• 41 Bini A, Fenoglio Jr J, Sobel J, Owen J, Fejgl M, Kaplan KL. Immunochemical characterization of fibrinogen, fibrin I and fibrin II in human thrombi and atherosclerotic lesions. Blood 1987; 69: 1038-1045