Heparin Regulates ICAM-1 Expression in Human Endothelial Cells: An Example of Non-Cytokine-Mediated Endothelial Activation

Summary

Activated endothelial cells up-regulate the expression of several molecules on their plasma membranes, including intercellular adhesion molecule-1 (ICAM-1). The role of heparin in regulating endothelial cell gene expression is unclear. We thus have investigated the ability of heparin to regulate ICAM-1 gene expression by using flow cytometry and the ribonuclease protection assay with human umbilical vein and aortic endothelial cells cultured in growth medium supplemented with 90 μg/ml heparin (heparin-sufficient, HS) or in growth medium without added heparin (heparin-deficient, HD). We found that HD medium increased plasma membrane protein and mRNA for ICAM-1 but not for HLA-DR, even though both ICAM-1 and HLA-DR protein and mRNA were inducible by gamma interferon (IFN-γ). In addition, phorbol ester and IFN-γ increased the expression of plasma membrane ICAM-1 or ICAM-1 and HLA-DR, respectively, more in HD medium than in HS medium. We found that the HD-mediated increase of ICAM-1 mRNA was reversible by the addition of heparin, and that the half-life of ICAM-1 mRNA was the same in both HS- and HD-treated cells. Also, heparin was found to suppress increases in ICAM-1 mRNA at a concentration as low as 5 μg/ml. These findings indicate that heparin deficiency induces endothelial activation characterized by increased ICAM-1, and that such induction is not dependent on cytokines or endotoxin. The modulation of ICAM-1 expression by heparin appears to occur at the transcriptional level. Thus, heparin may have a role in regulating endothelial function by affecting the expression of ICAM-1, thereby impacting upon the trans-endothelial trafficking of leukocytes.

• References

• 1 Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: The multistep paradigm. Cell 1994; 76: 301-14.
• 2 Pober JS. Cytokine-mediated activation of vascular endothelium. Am J Pathol 1988; 133: 426-33.
• 3 Cotran RS, Pober JS. Endothelial activation: Its role in inflammatory and immune reactions. In: Endothelial Cell Biology in Health and Disease. Simionescu N, Simionescu M. eds. New York, NY: Plenum Press; 1988: 335-47.
• 4 Makgoba MW, Sanders ME, Ginther Luce GE, Dustin ML, Springer TA, Clark EA, Mannoni P, Shaw S. ICAM-1 is a ligand for LFA-1-dependent adhesion of B, T and myeloid cells. Nature 1988; 331: 86-8.
• 5 Marlin SD, Springer TA. Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen-1 (LFA-1). Cell 1987; 51: 813-9.
• 6 Diamond MS, Staunton DE, de Fougerolles AR, Stacker SA, Garcia-Aguilar J, Hibbs M, Springer TA. ICAM-1 (CD54): A counter-receptor for Mac-1 (CD11b/CD18). J Cell Biol 1990; 111: 3129-39.
• 7 Dustin ML, Rothlein R, Bhan AK, Dinarello CA, Springer TA. Induction by IL 1 and interferon-gamma: Tissue distribution, biochemistry, and function of a natural adherence molecule (ICAM-1). J Immunol 1986; 137: 245-54.
• 8 Rothlein R, Czajkowski M, O’Neill MM, Marlin SD, Mainolfi E, Merluzzi VJ. Induction of intercellular adhesion molecule 1 on primary and continuous cell lines by pro-inflammatory cytokines. J Immunol 1988; 141: 1665-9.
• 9 Swerlick R, Garcia-Gonzalez Kubota Y, Xu Y, Lawley T. Studies of the modulation of the MHC antigen and cell adhesion molecule expression on human dermal microvascular endothelial cells. J Invest Dermatol 1991; 97: 190-6.
• 10 Cavender DE. Interactions between endothelial cells and the cells of the immune system. Int Rev Exp Pathol 1991; 32: 57-94.
• 11 Kaufman JF, Auffray C, Shackelford DA, Strominger J. The class II molecules of the human and murine major histocompatibility complex. Cell 1984; 36: 1-13.
• 12 Altmann DM, Hogg N, Trowsdale J, Wilkinson D. Cotransfection of ICAM-1 and HLA-DR reconstitutes human antigen-presenting cell function in mouse L cells. Nature 1989; 338: 512-4.
• 13 Bevilacqua MP, Nelson RM, Mannori G, Cecconi O. Endothelial-leukocyte adhesion molecules in human disease. Annu Rev Med 1994; 45: 361-78.
• 14 Poston RN, Haskard DO, Coucher JR, Gall NP, Johnson-Tidey RR. Expression of intercellular adhesion molecule-1 in atherosclerotic plaques. Am J Pathol 1992; 140: 665-73.
• 15 van der Wal AC, Das PK, Tigges AJ, Becker AE. Adhesion molecules on the endothelium and mononuclear cells in human atherosclerotic lesions. Am J Pathol 1992; 141: 1427-33.
• 16 Labarrere CA, Pitts D, Nelson DR, Faulk WP. Coronary artery disease in cardiac allografts: Association with arteriolar endothelial HLA-DR and ICAM-1 antigens. Transplant Proc 1995; 27: 1939-40.
• 17 Salomon RN, Hughes CCW, Schoen FJ, Payne DD, Pober JS, Libby P. Human coronary transplant-associated artheriosclerosis. Am J Pathol 1991; 138: 791-8.
• 18 Labarrere CA, Nelson DR, Faulk WP. Endothelial activation and development of coronary artery disease in transplanted human hearts. J Am Med Assoc 1997; 278: 1169-75.
• 19 Arfors KE, Ley K. Sulfated polysaccharides in inflammation. J Lab Clin Med 1993; 121: 201-2.
• 20 Sy MS, Schneeberger E, McCluskey R, Greene MI, Rosenberg RD, Benacerraf B. Inhibition of delayed-type hypersensitivity by heparin depleted of anticoagulant activity. Cell Immunol 1983; 82: 23-32.
• 21 Frieri M, Metcalfe DD. Analysis of the effect of mast cell granules on lymphocyte blastogenesis in the absence and presence of mitogens: Identification of heparin as a granule-associated suppressor factor. J Immunol 1983; 131: 1942-8.
• 22 Weerwind PW, Maessen JG, van Tits LJ, Stad RK, Fransen EJ, de Jong DS, Penn OC. Influence of duraflo II heparin-treated extracorporeal circuits on the systemic inflammatory response in patients having coronary bypass. J Thorac Cardiovasc Surg 1995; 110: 1633-41.
• 23 Rogers C, Welt FGP, Karnovsky MJ, Edelman ER. Monocyte recruitment and neointimal hyperplasia in rabbits. Coupled inhibitory effects of heparin. Arterioscler Thromb Vasc Biol 1996; 16: 1312-8.
• 24 Nelson RM, Cecconi O, Roberts WG, Aruffo A, Linhardt RJ, Bevilacqua MP. Heparin oligosaccharides bind L- and P-selectin and inhibit acute inflammation. Blood 1993; 82: 3253-8.
• 25 Tangelder GJ, Arfors KE. Inhibition of leukocyte rolling in venules by protamine and sulfated polysaccharides. Blood 1991; 77: 1565-71.
• 26 Diamond MS, Alon R, Parkos CA, Quinn MT, Springer TA. Heparin is an adhesive ligand for the leukocyte integrin Mac-1 (CD11b/CD18). J Cell Biol 1995; 130: 1473-82.
• 27 Korlipara LVP, Pleass HCC, Taylor RMR, Proud G, Forsythe JLR, Kirby JA. Heparin antagonizes the induction of class II MHC molecules by inter-feron-γ. Transplantation 1994; 58: 1426-9.
• 28 Rix DA, Douglas MS, Talbot D, Dark JH, Kirby JA. Role of glycosaminoglycans (GAGs) in regulation of the immunogenicity of human vascular endothelial cells. Clin Exp Immunol 1996; 104: 60-5.
• 29 Lantz M, Thysell H, Nilsson E, Olsson I. On the binding of tumor necrosis factor (TNF) to heparin and the release in vivo of the TNF-binding protein I by heparin. J Clin Invest 1991; 88: 2026-31.
• 30 Lortat-Jacob H, Grimaud JA. Binding of interferon-gamma to heparan sulfate is restricted to the heparin-like domains and involves carboxylicbut not N-sulfated-groups. Biochim Biophys Acta 1992; 1117: 126-30.
• 31 Lortat-Jacob H, Kleinman HK, Grimaud JA. High-affinity binding of inter-feron-γ to a basement membrane complex (Matrigel). J Clin Invest 1991; 87: 878-83.
• 32 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical vein: Identification by morphological and immunologic criteria. J Clin Invest 1973; 52: 2745-56.
• 33 Maciag T, Cerundolo J, Ilsley S, Kelley PR, Forand R. An endothelial cell growth factor from bovine hypothalamus: Identification and partial characterization. Proc Natl Acad Sci USA 1979; 76: 5674-8.
• 34 Justus AC, Roussev R, Norcross JL, Faulk WP. Antithrombin binding by human umbilical vein endothelial cells: Effects of exogenous heparin. Thromb Res 1995; 79: 175-86.
• 35 Yeh CG, Hsi B, Faulk WP. Propidium iodide as a nuclear marker in immunofluorescence. II. Use with cellular identification and viability studies. J Immunol Methods 1981; 43: 269-75.
• 36 Ambion.. Preparation and Purification of Radiolabeled RNA Probe. In: RPA II™ Ribonuclease Protection Assay Kit Instruction Manual. Austin, TX: Ambion, Inc.; 1993: 32-5.
• 37 Myers CL, Wertheimer SJ, Schembri-King J, Parks T, Wallace RW. Induction of ICAM-1 by TNF-α, IL-1β and LPS in human endothelial cells after downregulation of PKC. Am J Physiol 1992; 263: C767-C72.
• 38 Myers CL, Desai SN, Schembri-King J, Letts GL, Wallace RW. Discriminatory effects of protein kinase inhibitors and calcium ionophore on endothelial ICAM-1 induction. Am J Physiol 1992; 262: C365-C73.
• 39 Au YPT, Montgomery KF, Clowes AW. Heparin inhibits collagenase gene expression mediated by phorbol ester-responsive element in primate arterial smooth muscle cells. Circ Res 1992; 70: 1062-9.
• 40 Busch SJ, Martin GA, Barnhart RL, Mano M, Cardin AD. Trans-repressor activity of nuclear glycosaminoglycans on Fos and Jun/AP-1 oncoprotein-mediated transcription. J Cell Biol 1992; 116: 31-42.
• 41 Voraberger G, Schafer R, Stratowa C. Cloning of the human gene for intercellular adhesion molecule 1 and analysis of its 5’-regulatory region. J Immunol 1991; 147: 2777-86.
• 42 Karin M. The regulation of AP-1 activity by mitogen-activated protein kinases. J Biol Chem 1995; 270: 16483-6.
• 43 Angel P, Karin M. The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. Biochim Biophys Acta 1991; 1072: 129-57.
• 44 Au YPT, Dobrowolska G, Morris DR, Clowes AW. Heparin decreases activator protein-1 binding to DNA in part by posttranslational modification of Jun B. Circ Res 1994; 75: 15-22.
• 45 Degen SJF, Rajput B, Reich E. The human tissue plasminogen activator gene. J Biol Chem 1986; 261: 6972-85.
• 46 Au YPT, Kenagy RD, Clowes AW. Heparin selectively inhibits the transcription of tissue-type plasminogen activator in primate arterial smooth muscle cells during mitogenesis. J Biol Chem 1992; 267: 3438-44.
• 47 Collins T, Williams A, Johnston GI, Kim J, Eddy R, Shows T, Gimbrone MA, Bevilacqua MP. Structure and chromosomal location of the gene for endothelial-leukocyte adhesion molecule 1. J Biol Chem 1991; 266: 2466-73.
• 48 Pohlman TH, Harlan JM, Lobb R, Bomsztyk K, Tarr PI, Vassallo C, Goff D, Tizard R, Hession C, Osborn L, Montgomery KF. Activation of endothelial-leukocyte adhesion molecule 1 (ELAM-1) gene transcription. Proc Natl Acad Sci USA 1991; 88: 6523-7.
• 49 Jaques LB, Hiebert LM, Wice SM. Evidence from endothelium of gastric absorption of heparin and of dextran sulfate 8000. J Lab Clin Med 1991; 117: 122-30.
• 50 Vannucchi S, Pasquali F, Porciatti F, Chiarugi V, Magnelli L, Bianchini P. Binding, internalization and degradation of heparin and heparin fragments by cultured endothelial cells. Thromb Res 1988; 49: 373-83.
• 51 Barzu T, Molho P, Tobelem G, Petitou M, Caen J. Binding and endocytosis of heparin by human endothelial cells in culture. Biochim Biophys Acta 1985; 845: 196-203.
• 52 Fedarko NS, Conrad HE. A unique heparan sulfate in the nuclei of hepatocytes: Structural changes with the growth state of the cells. J Cell Biol 1986; 102: 587-99.
• 53 Au YPT, Kenagy RD, Clowes MM, Clowes AW. Mechanisms of inhibition by heparin of vascular smooth muscle cell proliferation and migration. Haemostasis 1993; 23 (Suppl. 01) 177-82.
• 54 Auwerx J, Sassone-Corsi P. IP-1: A dominant inhibitor of Fos/Jun whose activity is modulated by phosphorylation. Cell 1991; 64: 983-93.
• 55 Nader HB, Buonassisi V, Colburn P, Dietrich CP. Heparin stimulates the synthesis and modifies the sulfation pattern of heparan sulfate proteoglycan from endothelial cells. J Cell Physiol 1989; 140: 305-10.
• 56 Nader HB, Toma L, Pinhal MAS, Buonassisi V, Colburn P, Dietrich CP. Effect of heparin and dextran sulfate on the synthesis and structure of heparan sulfate from cultured endothelial cells. Semin Thromb Hemost 1991; 17: 47-56.
• 57 Gamble JR, Khew-Goodall Y, Vadas MA. Transforming growth factor-β inhibits E-selectin expression on human endothelial cells. J Immunol 1993; 150: 4494-503.
• 58 Kluszynski BA, Kim C, Faulk WP. Zinc as a cofactor for heparin neutralization by histidine-rich glycoprotein. J Biol Chem 1997; 272: 13541-7.
• 59 Volpi N, Cusmano M, Venturelli T. Qualitative and quantitative studies of heparin and chondroitin sulfates in normal human plasma. Biochim Biophys Acta 1995; 1243: 49-58.
• 60 Mazzoni MC, Schmid-Schonbein GW. Mechanisms and consequences of cell activation in the microcirculation. Cardiovasc Res 1996; 32: 709-19.