Evidence for the Expression of Urokinase-type Plasminogen Activator by Human Venous Endothelial Cells In Vivo

L. Camoin; R. Pannell; F. Anfosso; J. P. Lefevre; J. Sampol; V. Gurewich; F. Dignat-George

doi:10.1055/s-0037-1615396

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1998; 80(06): 961-967
DOI: 10.1055/s-0037-1615396

Letters to the Editor

Schattauer GmbH

Evidence for the Expression of Urokinase-type Plasminogen Activator by Human Venous Endothelial Cells In Vivo

L. Camoin

¹Laboratoire d’lmmuno-Hématologie, Faculté de Pharmacie

,

R. Pannell

³Vascular Research Laboratory, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

,

F. Anfosso

¹Laboratoire d’lmmuno-Hématologie, Faculté de Pharmacie

,

J. P. Lefevre

²Service d’Hématologie d’Hôpital de la Conception, Marseille, France

,

J. Sampol

¹Laboratoire d’lmmuno-Hématologie, Faculté de Pharmacie

,

V. Gurewich

³Vascular Research Laboratory, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

,

F. Dignat-George

¹Laboratoire d’lmmuno-Hématologie, Faculté de Pharmacie

› Author Affiliations

Abstract

Summary

Endothelial cells (ECs) in culture synthesize and secrete urokinase-type plasminogen activator (u-PA), but the normal vascular endothelium is believed to synthesize only tissue plasminogen activator (t-PA), which is thought to be responsible for intravascular fibrinolysis. More recently, animal studies have shown that the biological role of u-PA in fibrinolysis has been underestimated, prompting a re-examination of its synthesis by the endothelium. In this study, we investigated whether u-PA was synthesized by non-atherosclerotic endothelial cells in vivo by testing ECs dislodged by venipuncture from 12 normal volunteers and 17 patients admitted for plasmapheresis. The ECs were isolated with an anti-endothelial monoclonal antibody coupled to immunomagnetic beads and characterized by morphology and by labelling for vWF, CD31, and UEA-1 binding. U-PA antigen was found in 50% of the ECs from the normal subjects and in 60% of those from patients. U-PA enzymatic activity on zymograms was detected in 50% of the normal samples and 60% of the patient samples, with the latter being more frequently and more strongly positive. U-PA mRNA was found in all the normal and patient samples tested. The results indicate that u-PA is synthesized by the venous endothelium in vivo but that its expression is highly variable.

Full Text

References

References
1 Wun T-C, Capuano A. Spontaneous fibrinolysis in whole human plasma. Identification of tissue activator-related protein as the major plasminogen activator causing spontaneous activity in vitro . J Biol Chem 1985; 260: 5061-6.
2 Rijken DC, Wijngaards G, Welbergen J. Relationship between tissue plasminogen activator and the activators in blood and vascular wall. Thromb Res 1980; 18: 815-30.
3 Kristensen P, Larsson L-I, Nielsen LS, Grøndahl-Hansen J, Andreasen PA, Danø K. Human endothelial cells contain one type of plasminogen activator. FEBS Lett 1984; 168: 33-7.
4 Falkenberg M, Tjarnstrom J, Ortenwall P, Olausson M, Risberg B. Localization of fibrinolytic activators and inhibitors in normal and athero-sclerotic vessels. Thromb Haemost 1996; 75: 933-8.
5 Booyse FM, Scheinbuks J, Lin PH, Traylor M, Bruce R. Isolation and interrelationships of the multiple molecular tissue-type and urokinase-type plasminogen activator forms produced by cultured human umbilical vein endothelial cells. J Biol Chem 1988; 263: 15129-38.
6 Takahashi K, Kiguchi T, Sawasaki Y, Karikusa F, Nemoto N, Matsuoka T, Yamamoto M. Lung capillary endothelial cells produce and secrete urokinase-type plasminogen activator. Am J Respir Cell Mol Biol 1992; 7: 90-4.
7 van Hinsbergh VWM, Binnema D, Scheffer MA, Sprengers ED, Kooistra T, Rijken C. Production of plasminogen activators and inhibitor by serially propagated endothelial cells from adult human blood vessels. Arteriosclerosis 1987; 7: 389-400.
8 Wojta J, Hoover RL, Daniel TO. Vascular origin determines plasminogen activator expression in human endothelial cells. Renal endothelial cells produce large amounts of single chain urokinase type plasminogen activator. J Biol Chem 1989; 264: 2846-52.
9 Pepper MS, Sappino A-P, Stocklin R, Montesano R, Orci L, Vassalli J-D. Upregulation of urokinase receptor expression on migrating endothelial cells. J Cell Biol 1993; 122: 673-84.
10 Carmeliet P, Schoonjans L, Kieckens L, Ream B, Degen J, Bronson R, De Vos R, van den Oord JJ, Collen D, Mulligan RC. Physiological consequences of loss of plasminogen activator gene function in mice. Nature 1994; 368: 419-24.
11 Yamamoto K, Loskutoff DJ. Fibrin deposition in tissues from endotoxin-treated mice correlates with decreases in the expression of urokinase-type but not tissue-type plasminogen activator. J Clin Invest 1996; 97: 2440-51.
12 Huisveld IA, Kluft C, Hospers AJH, Bernik MJE, Erich WBM, Bouma BN. Effect of exercise and oral contraceptive agents on fibrinolytic potential in trained females. J Appl Physiol 1984; 56: 906-12.
13 Dooijewaard G, de Boer A, Turion PNC, Cohen AF, Breimer DD, Kluft C. Physical exercise induces enhancement of urokinase-type plasminogen activator (u-PA) levels in plasma. Thromb Haemost 1991; 65: 82-6.
14 Potter van Loon BJ, Heere LP, Kluft C, Briet E, Dooijewaard G, Meinders AE. Fibrinolytic system during long-distance running in IDDM patients and in healthy subjects. Diabetes Care 1992; 15: 991-8.
15 George F, Brisson C, Poncelet P, Laurent JC, Massot O, Arnoux D, Ambrosi P, Klein-Soyer C, Cazenave JP, Sampol J. Rapid isolation of human endothelial cells from whole blood using s-endo1 monoclonal antibody coupled to immuno-magnetic beads: demonstration of endothelial injury after angioplasty. Thromb Haemost 1992; 67: 147-53.
16 George F, Brouqui P, Boffa MC, Mutin M, Drancourt J, Brisson C, Raoult D, Sampol J. Demonstration of Rickettsia Conorii induced endothelial injury in vivo by measuring circulating endothelial cells, thrombomodulin and von Willebrand factor in patients with Mediterranean Spotted Fever. Blood 1993; 82: 2109-16.
17 Lijazi A, Devonec M, Bouvier R, Escourrou G, Longin A, Perrin P, Revillard VP. Phenotyping of 76 human bladder tumors with a panel of monoclonal antibodies. Correlation between pathology, surface immunofluorescence and DNA content. Eur J Clin Oncol 1982; 25: 777-85.
18 Jeanneau C, Sultan Y. Localization of factor VIII/von Willebrand factor antigen by immunoelectron microscopy in human endothelial cells using Fab fragments coupled to peroxidase. J Histochem Cytochem 1982; 30: 1091-9.
19 Brochier J, Abou Hamed A, Gudro JP, Revillard JP. Study of human T and B lymphocytes with heterologous antisera. Preparation specificity and properties of antisera. Immunology 1976; 31: 749-57.
20 Chamley-Campbell JH, Campbell GR, Ross R. Phenotype dependent response of cultured aortic smooth muscle to serum mitogens. J Cell Biol 1981; 89: 379-83.
21 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5.
22 Krieg PA, Melton DA. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nuclei Acid Research 1984; 12: 7057-64.
23 Wilcox JN. Fundamental principles of in situ hybridization. J Histochem Cytochem 1993; 41: 1725-33.
24 Nielsen LS, Kellerman GM, Behrendt N, Picone R, Dano K, Blasi F. A 55,000-60,000 Mr receptor protein for urokinase-type plasminogen activator. Identification in human tumor cell lines and partial purification. J Biol Chem 1988; 263: 2358-63.
25 Northeast ADR, Soo KS, Bobrow LG, Gaffney PJ, Burnand KG. The tissue plasminogen activator and urokinase response in vivo during natural resolution of venous thrombus. J Vasc Surg 1995; 22: 573-9.
26 Pannell R, Black J, Gurewich V. The complementary modes of action of tissue plasminogen activator (t-PA) and pro-urokinase (pro-UK) by which their synergistic effect on clot lysis may be explained. J Clin Invest 1988; 81: 853-9.
27 Liu J, Gurewich V. A comparative study of the promotion of tissue plasminogen activator and pro-urokinase-induced plasminogen activation by fragments D and E-2 of fibrin. J Clin Invest 1991; 88: 2012-7.
28 Larsson L-I, Skriver L, Nielsen LS, Grøndahl-Hansen J, Kristensen P, Danø K. Distribution of urokinase-type plasminogen activator immunoreactivity in the mouse. J Cell Biol 1984; 98: 894-903.
29 van Hinsbergh VWM, Kooistra T, Emeis JJ, Koolwijk P. Regulation of plasminogen activator production by endothelial cells: role in fibrinolysis and local proteolysis. Int J Radiat Biol 1991; 60: 261-8.
30 Lupu F, Heim DA, Bachmann F, Hurni M, Kakkar VV, Kruithof EKO. Plasminogen activator expression in human atherosclerotic lesions. Arterioscler Thromb Vasc Biol 1995; 15: 1444-55.
31 Padro T, Emeis JJ, Steins M, Schmid KW, Kienast J. Quantification of plasminogen activators and their inhibitors in the aortic vessel wall in relation to the presence and severity of atherosclerotic disease. Arterioscler Thromb Vasc Biol 1995; 15: 893-902.
32 Schneiderman J, Bordin GM, Engelberg I, Adar R, Seiffert D, Thinnes T, Bernstein EF, Dilley RB, Loskutoff D. Expression of fibrinolytic genes in atherosclerotic abdominal aortic aneurysm wall. J Clin Invest 1995; 96: 639-45.
33 Raghunath PN, Tomaszewski JE, Brady ST, Caron RJ, Okada SS, Barna-than ES. Plasminogen activator system in human coronary atherosclerosis. Arterioscler Thromb Vasc Biol 1995; 15: 1432-43.
34 Kienast J, Padro T, Steins M, Li CX, Schmid KW, Hammel D, Scheld HH, van de Loo JCW. Relation of urokinase-type plasminogen activator expression to presence and severity of atherosclerotic lesions in human coronary arteries. Thromb Haemost 1998; 79: 579-86.
35 Kjaeldgaard A, Larsson B, Astedt B. Release of both urokinase and tissue plasminogen activator from veins in vitro . Thromb Res 1986; 44: 729-37.
36 Wojta J, Binder BR, Huber K, Hoover RL. Evaluation of fibrinolytic capacity in plasma during thrombolytic therapy with single (scu-PA) or two-chain urokinase type plasminogen activator (tcu-PA) by a combined assay system for urokinase-type plasminogen activator antigen and function. Thromb Haemost 1989; 61: 289-93.
37 Behrendt N, Rønne E, Danø K. The structure and function of the urokinase receptor, a membrane protein governing plasminogen activation on the cell surface. Biol Chem Hoppe-Seyler 1995; 376: 269-79.
38 Sato Y, Rifkin DB. Autocrine activities of basic fibroblast growth factor: regulation of endothelial cell movement, plasminogen activator synthesis and DNA synthesis. J Cell Biol 1988; 107: 1199-206.
39 van Hinsbergh VWM, van den Berg EA, Fiers W, Dooijewaard G. Tumor necrosis factor induces the production of urokinase-type plasminogen activator by human endothelial cells. Blood 1990; 75: 1991-8.
40 Lengyel E, Klostergaard J, Boyd D. Stimulation of urokinase expression by TNF-α requires the activation of binding sites for the AP-1 and PEA3 transcription factors. Biochim Biophys Acta 1995; 1268: 65-72.
41 Mandriota SJ, Seghezzi G, Vassalli J-D, Ferrara N, Wasi S, Mazzieri R, Mignatti P, Pepper MS. Vascular endothelial growth factor increases urokinase receptor expression in vascular endothelial cells. J Biol Chem 1995; 270: 9709-16.
42 Li C, Jiang Y, Tang W, Gurewich V, Liu JN. Auto-induction of urokinase-type plasminogen activator (u-PA) in endothelial cells and monocytes. Thromb Haemost. XVth Congr ISTH 77: PS-3070, 1997 (abstr).