Proinflammatory properties of murine aortic endothelial cells exclusively expressing a non cleavable form of TNFα

Summary

Soluble (sTNF) and transmembrane (tmTNF) forms of TNFα (TNF) have distinct proinflammatory effects. We investigated whether tmTNF altered the synthesis of some proinflammatory proteins involved in atherothrombosis, in murine aortas and aortic endothelial cells (MAEC). Samples were obtained from wild-type (WT) mice and TNF-deficient mice that express a mutated non cleavable tmTNF transgene (tmTNFnc).The levels of secreted MCP-1, RANTES, IL-6, PAI-1, soluble ICAM-1, and soluble TNF receptor type 1 (TNFR1; CD120a) antigens, MMP-9 activity and of cell surface ICAM-1 were not significantly different between the two types of MAEC.The magnitude of endotoxin-stimulated production of RANTES, MCP-1 and IL-6 was similar in the two types of cells. Of note, the amount of synthesized TNF receptor type 2 (TNFR2; CD120b), measured by its secreted (in aorta and MAEC), intracellular and mRNA levels (in MAEC), was significantly 4-fold lower in tmTNFnc than in WT mice, both in basal and endotoxin-stimulated conditions. A neutralizing anti-TNF antibody or the recombinant murine TNF did not modify the magnitude of the difference in TNFR2 production between the two types of cells, suggesting a preponderant role of tmTNF in the down-regulation of TNFR2 synthesis. Macrophages of tmTNFnc mice also produced less TNFR2 than WT macrophages (−30%). Plasmas of tmTNFnc mice contained significantly less sTNFR2 than WT mice (−75%). In conclusion, an increase in tmTNF levels, rather than the lack of sTNF, significantly down-modulated TNFR2 synthesis in aortic endothelial cells, but had no major influence on the synthesis of some major pro-inflammatory and pro-atherothrombotic proteins.

Keywords

Transmembrane TNFα - TNFR2/CD120b - aortic endothelial cells - cytokines - chemokines

• References

• 1 Moss ML, Jin SL, Milla ME. et al. Cloning of a disintegrin metalloproteinase that processes precursor tumour-necrosis factor-alpha. Nature 1997; l385: 733-6.
• 2 Black RA, Rauch CT, Kozlosky CJ. et al. A metalloproteinase disintegrin that releases tumour-necrosis factor- alpha from cells. Nature 1997; 385: 729-33.
• 3 Peschon JJ, Slack JL, Reddy P. et al. An essential role for ectodomain shedding in mammalian development. Science 1998; 282: 1281-4.
• 4 Reddy P, Slack JL, Davis R. et al. Functional analysis of the domain structure of tumor necrosis factor- alpha converting enzyme. J Biol Chem 2000; 275: 14608-14.
• 5 Solomon KA, Pesti N, Wu G. et al. Cutting edge: a dominant negative form of TNF-alpha converting enzyme inhibits proTNF and TNFRII secretion. J Immunol 1999; 163: 4105-8.
• 6 Aderka D, Engelmann H, Maor Y. et al. Stabilization of the bioactivity of tumor necrosis factor by its soluble receptors. J Exp Med 1992; 175: 323-9.
• 7 Terlizzese M, Simoni P, Antonetti F. In vitro comparison of inhibiting ability of soluble TNF receptor p75 (TBP II) vs soluble TNF receptor p55 (TBP I) against TNF-alpha and TNF-beta. J Interferon Cytokine Res 1996; 16: 1047-53.
• 8 Peiretti F, Bernot D, Lopez S. et al. Modulation of PAI-1 and proMMP-9 syntheses by soluble TNFalpha and its receptors during differentiation of the human monocytic HL-60 cell line. J Cell Physiol 2003; 196: 346-53.
• 9 Kriegler M, Perez C, DeFay K. et al. A novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell 1988; 53: 45-53.
• 10 Perez C, Albert I, DeFay K. et al. A nonsecretable cell surface mutant of tumor necrosis factor (TNF) kills by cell-to-cell contact. Cell 1990; 63: 251-8.
• 11 Haas E, Grell M, Wajant H. et al. Continuous autotropic signaling by membrane-expressed tumor necrosis factor. J Biol Chem 1999; 274: 18107-12.
• 12 Grell M, Zimmermann G, Gottfried E. et al. Induction of cell death by tumour necrosis factor (TNF) receptor 2, CD40 and CD30: a role for TNF-R1 activation by endogenous membraneanchored TNF. EMBO J 1999; 18: 3034-43.
• 13 Caron G, Delneste Y, Aubry JP. et al. Human NK cells constitutively express membrane TNF-alpha (mTNFalpha) and present mTNFalpha-dependent cytotoxic activity. Eur J Immunol 1999; 29: 3588-95.
• 14 Eissner G, Kohlhuber F, Grell M. et al. Critical involvement of transmembrane tumor necrosis factor-alpha in endothelial programmed cell death mediated by ionizing radiation and bacterial endotoxin. Blood 1995; 86: 4184-93.
• 15 Decoster E, Vanhaesebroeck B, Boone E. et al. Induction of unresponsiveness to tumor necrosis factor (TNF) after autocrine TNF expression requires TNF membrane retention. J Biol Chem 1998; 273: 3271-7.
• 16 Eissner G, Kirchner S, Lindner H. et al. Reverse signaling through transmembrane TNF confers resistance to lipopolysaccharide in human monocytes and macrophages. J Immunol 2000; 164: 6193-8.
• 17 Tanabe Y, Kohchi C, Kitahara-Tanabe N. et al. Involvement of 26-kDa membrane-bound tumor necrosis factor precursor in bidirectional feedback regulation on 17-kDa tumor necrosis factor production after stimulation by lipolysaccharide. Cytokine 1998; 10: 82-92.
• 18 Lindner H, Holler E, Ertl B. et al. Peripheral blood mononuclear cells induce programmed cell death in human endothelial cells and may prevent repair: Role of cytokines. Blood 1997; 89: 1931-8.
• 19 Clauss M, Sunderkotter C, Sveinbjornsson B. et al. A permissive role for tumor necrosis factor in vascular endothelial growth factorinduced vascular permeability. Blood 2001; 97: 1321-9.
• 20 Fujii T, Okada M, Mimori T. et al. The transmembrane form of TNFalpha drives autoantibody production in the absence of CD154: studies using MRL/Mp-Faslpr mice. Clin Exp Immunol 2002; 130: 224-32.
• 21 Grell M, Douni E, Wajant H. et al. The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor. Cell 1995; 83: 793-802.
• 22 Mueller C, Corazza N, Trachsel-Loseth S. et al. Noncleavable transmembrane mouse tumor necrosis factor-alpha (TNFalpha) mediates effects distinct from those of wild-type TNFalpha in vitro and in vivo . J Biol Chem 1999; 274: 38112-8.
• 23 Olleros ML, Guler R, Corazza N. et al. Transmembrane TNF induces an efficient cellmediated immunity and resistance to mycobacterium bovis bacillus Calmette-Guerin infection in the absence of secreted TNF and lymphotoxin-{alpha}. J Immunol 2002; 168: 3394-401.
• 24 Ruuls SR, Hoek RM, Ngo VN. et al. Membrane-bound TNF supports secondary lymphoid organ structure but is subservient to secreted TNF in driving autoimmune inflammation. Immunity 2001; 15: 533-43.
• 25 Willuweit A, Sass G, Schoneberg A. et al. Chronic inflammation and protection from acute hepatitis in transgenic mice expressing TNF in endothelial cells. J Immunol 2001; 167: 3944-52.
• 26 Canault M, Peiretti F, Mueller C. et al. Exclusive expression of transmembrane TNF-[alpha] in mice reduces the inflammatory response in early lipid lesions of aortic sinus*1. Atherosclerosis 2004; 172: 211-8.
• 27 Blankenberg S, Barbaux S, Tiret L. Adhesion molecules and atherosclerosis. Atherosclerosis 2003; 170: 191-203.
• 28 Sattar N. Inflammation and endothelial dysfunction: intimate companions in the pathogenesis of vascular disease?. Clin Sci (Lond) 2004; 106: 443-5.
• 29 Eugster HP, Muller M, Karrer U. et al. Multiple immune abnormalities in tumor necrosis factor and lymphotoxin-alpha double-deficient mice. Int Immunol 1996; 08: 23-36.
• 30 Kreisel D, Krupnick AS, Szeto WY. et al. A simple method for culturing mouse vascular endothelium. J Immunol Methods 2001; 254: 31-45.
• 31 Marelli-Berg FM, Peek E, Lidington EA. et al. Isolation of endothelial cells from murine tissue. J Immunol Methods 2000; 244: 205-15.
• 32 Declerck PJ, Verstreken M, Collen D. Immunoassay of murine t-PA, u-PA and PAI-1 using monoclonal antibodies raised in geneinactivated mice. Thromb Haemost 1995; 74: 1305-9.
• 33 Gerspach J, Gotz A, Zimmermann G. et al. Detection of membrane-bound tumor necrosis factor (TNF): an analysis of TNF-specific reagents. Microsc Res Tech 2000; 50: 243-50.
• 34 Ranta V, Orpana A, Carpen O. et al. Human vascular endothelial cells produce tumor necrosis factor-alpha in response to proinflammatory cytokine stimulation. Crit Care Med 1999; 27: 2184-7.
• 35 Imaizumi T, Itaya H, Fujita K. et al. Expression of tumor necrosis factor-{alpha} in cultured human endothelial cells stimulated with lipopolysaccharide or interleukin-1{alpha}. Arterioscler Thromb Vasc Biol 2000; 20: 410-5.
• 36 Beck G, Bottomley G, Bradshaw D. et al. (E)-2(R)-[1(S)-(Hydroxycarbamoyl)-4-phenyl-3-butenyl]-2’-isobutyl-2’-(methanesulfonyl)-4-methylvalerohydrazide (Ro 32-7315), a selective and orally active inhibitor of tumor necrosis factor-alpha convertase. J Pharmacol Exp Ther 2002; 302: 390-6.
• 37 Gallea-Robache S, Morand V, Millet S. et al. A metalloproteinase inhibitor blocks the shedding of soluble cytokine receptors and processing of transmembrane cytokine precursors in human monocytic cells. Cytokine 1997; 09: 340-6.
• 38 Slowik MR, De Luca LG, Fiers W. et al. Tumor necrosis factor activates human endothelial cells through the p55 tumor necrosis factor receptor but the p75 receptor contributes to activation at low tumor necrosis factor concentration. Am J Pathol 1993; 143: 1724-30.
• 39 Solomon KA, Pesti N, Wu G. et al. Cutting edge: A dominant negative form of TNF-{alpha} converting enzyme inhibits ProTNF and TNFRII secretion. J Immunol 1999; 163: 4105-8.
• 40 Bradley JR, Thiru S, Pober JS. Disparate localization of 55-kd and 75-kd tumor necrosis factor receptors in human endothelial cells. Am J Pathol 1995; 146: 27-32.
• 41 Harashima S-I, Horiuchi T, Hatta N. et al. Outside-to-inside signal through the membrane TNF-{{alpha}} induces E-selectin (CD62E) expression on activated human CD4+ T Cells. J Immunol 2001; 166: 130-6.
• 42 Kirchner S, Boldt S, Kolch W. et al. LPS resistance in monocytic cells caused by reverse signaling through transmembrane TNF (mTNF) is mediated by the MAPK/ERK pathway. J Leukoc Biol 2004; 75: 324-31.
• 43 Pocsik E, Duda E, Wallach D. Phosphorylation of the 26 kDa TNF precursor in mono-cytic cells and in transfected HeLa cells. J Inflamm 1995; 45: 152-60.
• 44 Watts AD, Hunt NH, Wanigasekara Y. et al. A casein kinase I motif present in the cytoplasmic domain of members of the tumour necrosis factor ligand family is implicated in `reverse signalling’. EMBO J 1999; 18: 2119-26.
• 45 Akassoglou K, Douni E, Bauer J. et al. Exclusive tumor necrosis factor (TNF) signaling by the p75TNF receptor triggers inflammatory ischemia in the CNS of transgenic mice. PNAS 2003; 100: 709-14.
• 46 Lucas R, Juillard P, Decoster E. et al. Crucial role of tumor necrosis factor (TNF) receptor 2 and membrane-bound TNF in experimental cerebral malaria. Eur J Immunol 1997; 27: 1719-25.
• 47 Porsch-Oezcueruemez M, Kunz D, Kloer H-U. et al. Evaluation of serum levels of solubilized adhesion molecules and cytokine receptors in coronary heart disease. J Am Coll Cardiol 1999; 34: 1995-2001.
• 48 Benjafield AV, Wang XL, Morris BJ. Tumor necrosis factor receptor 2 gene (TNFRSF1B) in genetic basis of coronary artery disease. J Mol Med 2001; 79: 109-15.
• 49 Douni E, Kollias G. A Critical role of the p75 tumor necrosis factor receptor (p75TNF-R) in organ inflammation independent of TNF, Lymphotoxin alpha , or the p55TNF-R. J Exp Med 1998; 188: 1343-52.