Protease Dependent Activation of Endothelial Cells by Peritoneal Dialysis Effluents

Summary

Incubation of cultured human umbilical vein endothelial cells (HUVECs) with dilutions of peritoneal dialysis effluents (PDEs) from 11 individual patients undergoing continuous ambulatory peritoneal dialysis (CAPD) induced cellular procoagulant activity in a dose and time dependent manner. This procoagulant activity could be attributed to tissue factor (TF) expression since it was blocked by rabbit anti-TF IgG. These data was confirmed by FACS analysis yielding surface TF expression; In addition PDEs induced the expression of E-selectin in HUVECs. This TF and selectin inducing activity was heat labile and could be inhibited by protease inhibitors. Partial purification could be achieved using a benzamidine-Sepharose column. The TF inducing activity could not be attributed to LPS, IL-1, TNF-α, mast cell tryptase, active thrombin, or complement factor D. We therefore conclude that the peritoneal cavity contains a protease activity that induces a procoagulatory and proinflammatory phenotype in HUVECs.

Key Words

Peritoneal dialysis - endothelial cells - tissue factor - adhesion molecules

• References

• 1 Nolph K, Lindbald AS, Novak JW. Continuos ambulatory peritoneal dialysis. N Engl J Med 1988; 318: 1595-600.
• 2 Pollock CA, Ibels LS, Eckstein RP, Graham JC, Caterson RJ, Mahony JF, Sheil AGR. Peritoneal morphology on maintainance dialysis. Am J Nephrol 1989; 9: 198-204.
• 3 Medcalf JF, Walls J. New frontiers in continuous ambulatory peritoneal dialysis. Kidney Int 1997; 52: S-108-10.
• 4 Goedde M, Sitter T, Schiffl H, Bechtel U, Schramm W, Spannagl M. Coagulation and fibrinolysis-related antigens in plasma and dialysate of CAPD patients. Perit Dial Int 1997; 17: 162-6.
• 5 Sitter T, Spannagl M, Schiffl H, Held E, van Hinsbergh VWM, Kooistra T. Imbalance between intraperitoneal coagulation and fibrinolysis during peritonitis of CAPD patients: the role of mesothelial cells. Nephrol Dial Transplant 1995; 10: 677-83.
• 6 Dobbie JW. Serositis: comparative analysis of histological findings and pathogenetic mechanisms in nonbacterial serosal inflammation. Perit Dial Int 1993; 13: 256-69.
• 7 Honda K, Nitta K, Horita S, Yumura W, Nihei H. Morphological changes in the peritoneal vasculature of patients on CAPD with ultrafiltration failure. Nephron 1997; 72: 171-6.
• 8 Osterud B. Tissue factor: a complex biological role. Thromb Haemost 1997; 78: 755-8.
• 9 Moll T, Czyz M, Holzmüller H, Hofer-Warbinek R, Wagner E, Winkler H, Bach FH, Hofer E. Regulation of the tissue factor promoter in endothelial cells. J Biol Chem 1995; 270: 3849-67.
• 10 Nawroth PP, Stern DM. Modulation of endothelial cell hemostatic properties by tumor necrosis factor. J Exp Med 1986; 163: 740-5.
• 11 Bevilacqua MP, Pober JS, Majeau GR, Cotran RS, Gimbrone MA. Interleukin 1 (IL-1) induces biosynthesis and cell surface expression of procoagulant activity in human vascular endothelial cells. J Exp Med 1984; 160: 618-23.
• 12 Moore KL, Andreoli SP, Esmon NL, Esmon CT, Bang NU. Endotoxin enhances tissue factor and suppresses thrombomodulin expression of human vascular endothelium in vitro. J Clin Invest 1987; 79: 124-30.
• 13 Tedesco F, Pausa M, Nardon E, Introna M, Mantovani A, Dobrina A. The cytolytically inactive terminal complement complex activates endothelial cells to express adhesion molecules and tissue factor procoagulant activity. J Exp Med 1997; 185: 1619-27.
• 14 Ikeda K, Nagasawa K, Horiuchi T, Tsuru T, Nishizaka H, Niho Y. C5a induces tissue factor activity on endothelial cells. Thromb Haemost 1997; 77: 394-8.
• 15 Clauss M, Gerlach M, Gerlach H, Brett J, Wang F, Familletti PC, Pan Y-CE, Olander JV, Conolly DT, Stern D. Vascular permeability factor: a tumor-derived polypeptide that induces endothelial cell and monocyte procoagulant activity, and promotes monocyte migration. J Exp Med 1990; 172: 1535-45.
• 16 Nishimura H, Tsuji H, Masuda H, Nakagawa K, Nakahara Y, Kitamura H, Kasahara T, Sugano T, Yoshizumi M, Sawada S, Nakagawa M. Angiotensin II increases plasminogen activator inhibitor-1 and tissue factor mRNA expression without changing that of tissue type plasminogen activator or tissue factor pathway inhibitor in cultured rat aortic endothelial cells. Thromb Haemost 1997; 77: 1189-95.
• 17 Bartha K, Brisson C, Archipoff G, de la Salle C, Lanza F, Cazenave JP, Beretz A. Thrombin regulates tissue factor and thrombomodulin mRNA levels and activities in human saphenous vein endothelial cells by distinct mechanisms. J Biol Chem 1993; 268: 421-9.
• 18 Ishihara H, Connolly AJ, Zeng D, Kahn ML, Zheng YW, Timmons C, Tram T, Coughlin SR. Protease-activated receptor 3 is a second thrombin receptor in humans. Nature 1997; 386: 502-6.
• 19 Molino M, Woolkalis MJ, Reavey-Cantwell J, Pratico D, Andrade-Gordon P, Barnathan ES, Brass LF. Endothelial cell thrombin receptors and PAR-2. J Biol Chem 1997; 272: 111333-41.
• 20 Wertheimer SJ, Myers CL, Wallace RW, Parks TP. Intercellular adhesion molecule-1 gene expression in human endothelial cells. J Biol Chem 1992; 267: 12030-5.
• 21 Osborn L, Hession C, Tizard R, Vasallo C, Luhowskyj S, Chi Rosso G, Lobb R. Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds to lymphocytes. Cell 1989; 59: 1203-11.
• 22 Kansas GS. Selectins and their ligands: current concepts and controversies. Blood 1996; 88: 3259-87.
• 23 Smith CW. Leukocyte-endothelial cell interactions. Semin Hematol 1993; 30 (04) (Suppl. 04) 45-55.
• 24 Shankar R, de la Motte CA, Poptic EJ, DiCorleto PE. Thrombin receptor-activating peptides differentially stimulate platelet-derived growth factor production, monocytic cell adhesion, and E-selectin expression in human umbilical vein endothelial cells. J Biol Chem 1994; 269: 13936-41.
• 25 Harboe N, Ingild A. Immunization, isolation of immunoglobulins, estimation of antibody titre. In: A manual of quantitative immunelectrophoresis. Methods and applications Axelsen NH, Kroll J, Weeke B. (eds). Universitetsforlaget Oslo; 1973: 161-46.
• 26 Zhang JC, Fabry A, Paucz L, Wojta J, Binder BR. Human fibroblasts down-regulate plasminogen activator inhibitor type-1 in cultured human macro-vascular and microvascular endothelial cells. Blood 1996; 88: 3880-6.
• 27 Shankar R, de la Motte CA, DiCorleto PE. 3-deazaadenosine inhibits thrombin-stimulated platelet-derived growth factor production and endothelial-leukocyte adhesion molecule-1-mediated monocytic cell adhesion in human aortic endothelial cells. J Biol Chem 1992; 267: 9376-82.
• 28 Winkler ME, Blaber M. Purification and characterization of recombinant single-chain urokinase produced in Escherichia coli. Biochemistry 1986; 25: 4041-5.
• 29 Mizuno K, Nakamura T, Takada K, Sakakibara S, Matsuo H. A membrane-bound, calcium-dependent protease in yeast α-cell cleaving on the carboxyl side of paired basic residues. Biochem Biophys Res Comm 1987; 144: 807-14.
• 30 Catana E, Schifferli JA. Purification of human complement factor D from the peritoneal fluid of patients on chronic ambulatory peritoneal dialysis. J Immunol Methods 1991; 138: 265-71.
• 31 Gardiner JE, Griffin H. Studies on human protein C inhibitor in normal and factor V/VIII deficient plasmas. Thromb Res 1984; 36: 197-203.
• 32 Molino M, Barnathan ES, Numerof R, Clark J, Dreyer M, Cumashi A, Hoxie JA, Schechter N, Woolkalis M, Brass LF. Interactions of mast cell tryptase with thrombin receptors and PAR-2. J Biol Chem 1997; 272: 4043-9.
• 33 Lewis S, Holmes C. Host defense mechanisms in the peritoneal cavity of continuous ambulatory peritoneal dialysis patients. Perit Dial Int 1991; 11: 14-21.
• 34 Brauner A, Hylander B, Wretlind B. Tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-1 receptor antagonist in dialysate and serum from patients on continuous ambulatory peritoneal dialysis. Am J Kidney Dis 1996; 27: 402-8.
• 35 Zemel D, Imholz ALT, Waart DR, Dinkla C, Struijk DG, Krediet RT. Appearance of tumor necrosis factor-α and soluble TNF-receptors I and II in peritoneal effluent of CAPD. Kidney Int 1994; 46: 1422-30.
• 36 van Hinsbergh VWM, Kooistra T, Scheffer MA, van Bockel JH, Goos NP, van Muijen NP. Characterization and fibrinolytic properties of human omental tissue mesothelial cells. Comparison with endothelial cells. Blood 1990; 75: 1490-7.
• 37 Totani L, Cumashi A, Piccoli A, Lorenzet R. Polymorphonuclear leukocytes induce PDGF release from IL-1α-treated endothelial cells. Role of adhesion molecules and serine proteases. Arterioscler Thromb Vasc Biol 1998; 18: 1534-40.