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Licensed Unlicensed Requires Authentication Published by De Gruyter September 6, 2005

Effects of elastase and cathepsin G on the levels of membrane and soluble TNFα

Renata Mężyk-Kopeć, Małgorzata Bzowska, Monika Bzowska, Barbara Mickowska, Paweł Mak, Jan Potempa and Joanna Bereta
From the journal

Abstract

Neutrophil elastase (NE) and cathepsin G (CG), the proteolytic enzymes localized in azurophil granules of neutrophils (PMN), are involved in PMN responses to various stimuli. When released at sites of inflammation, they participate in the degradation of numerous proteins involved in the regulation of the immune response. In this study, we employed ADAM17-/- fibroblasts stably transfected with cDNA of human pro-tumor necrosis factor α (proTNFα) (ADAM17-/-TNF+) to investigate the effects of NE and CG on shedding and degradation of TNFα. Both NE and CG were found to diminish the level of membrane TNFα (mTNFα) as measured by flow cytometry. This process was accompanied by the accumulation of biologically active soluble TNFα (sTNFα) in the culture medium, as determined by an increase in both the cytotoxic activity of TNFα and its ability to serve as a co-stimulator in the induction of inducible nitric oxide synthase (iNOS). However, in contrast to CG, NE at high concentrations was able to degrade sTNFα released from the cell surface. Using soluble recombinant human TNFα, we identified Val93-Ala94 and Val117-Glu118 as the NE cleavage sites within the sTNFα molecule. Taken together, the ability of NE and CG to modulate levels of membrane and soluble forms of TNFα may contribute to the proinflammatory activity of neutrophils.

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References

Aggarwal, B.B., Samanta, A., and Feldmann, M. (2001). TNFα. In: Cytokine Reference, Vol. I, S.K. Durum, T. Hirano, J. Vilcek and N.A. Nicola, eds. (London, UK: Academic Press), pp. 413–434.Search in Google Scholar

Ariel, A., Yavin, E.J., Hershkoviz, R., Avron, A., Franitza, S., Hardan, I., Cahalon, L., Fridkin, M., and Lider, O. (1998). IL-2 induces T cell adherence to extracellular matrix: inhibition of adherence and migration by IL-2 peptides generated by leukocyte elastase. J. Immunol.161, 2465–2472.Search in Google Scholar

Bank, U., Kupper, B., Reinhold, D., Hoffmann, T., and Ansorge, S. (1999a). Evidence for a crucial role of neutrophil-derived serine proteases in the inactivation of interleukin-6 at sites of inflammation. FEBS Lett.461, 235–240.10.1016/S0014-5793(99)01466-0Search in Google Scholar

Bank, U., Reinhold, D., Schneemilch, C., Kunz, D., Synowitz, H.J., and Ansorge, S. (1999b). Selective proteolytic cleavage of IL-2 receptor and IL-6 receptor ligand binding chains by neutrophil-derived serine proteases at foci of inflammation. J. Interferon Cytokine Res.19, 1277–1287.10.1089/107999099312957Search in Google Scholar

Bedard, M., McClure, C.D., Schiller, N.L., Francoeur, C., Cantin, A., and Denis, M. (1993). Release of interleukin-8, interleukin-6, and colony-stimulating factors by upper airway epithelial cells: implications for cystic fibrosis. Am. J. Respir. Cell Mol. Biol.9, 455–462.10.1165/ajrcmb/9.4.455Search in Google Scholar

Bereta, M., Bereta, J., Cohen, S., and Cohen, M.C. (1992). Low density lipoprotein inhibits accumulation of nitrites in murine brain endothelial cell cultures. Biochem. Biophys. Res. Commun.186, 315–320.10.1016/S0006-291X(05)80809-5Search in Google Scholar

Black, R.A., Rauch, C.T., Kozlosky, C.J., Peschon, J.J., Slack, J.L., Wolfson, M.F., Castner, B.J., Stocking, K.L., Reddy, P., Srinivasan, S. et al. (1997). A metalloproteinase disintegrin that releases tumour-necrosis factor-α from cells. Nature385, 729–733.10.1038/385729a0Search in Google Scholar

Bzowska, M., Guzik, K., Barczyk, K., Ernst, M., Flad, H.D., and Pryjma, J. (2002). Increased IL-10 production during spontaneous apoptosis of monocytes. Eur. J. Immunol.32, 2011–2020.10.1002/1521-4141(200207)32:7<2011::AID-IMMU2011>3.0.CO;2-LSearch in Google Scholar

Carden, D., Xiao, F., Moak, C., Willis, B.H., Robinson-Jackson, S., and Alexander, S. (1998). Neutrophil elastase promotes lung microvascular injury and proteolysis of endothelial cadherins. Am. J. Physiol.275, H385–392.10.1152/ajpheart.1998.275.2.H385Search in Google Scholar

Celis, J. (1998). Cell Biology: A Laboratory Handbook (London, UK: Academic Press), pp. 16–18.Search in Google Scholar

Chertov, O., Yang, D., Howard, O.M., and Oppenheim, J.J. (2000). Leukocyte granule proteins mobilize innate host defenses and adaptive immune responses. Immunol. Rev.177, 68–78.10.1034/j.1600-065X.2000.17702.xSearch in Google Scholar

Clauss, M., Sunderkotter, C., Sveinbjornsson, B., Hippenstiel, S., Willuweit, A., Marino, M., Haas, E., Seljelid, R., Scheurich, P., Suttorp, N., Grell, M., and Risau, W. (2001). A permissive role for tumor necrosis factor in vascular endothelial growth factor-induced vascular permeability. Blood97, 1321–1329.10.1182/blood.V97.5.1321Search in Google Scholar

Coeshott, C., Ohnemus, C., Pilyavskaya, A., Ross, S., Wieczorek, M., Kroona, H., Leimer, A.H., and Cheronis, J. (1999). Converting enzyme-independent release of tumor necrosis factor α and IL-1β from a stimulated human monocytic cell line in the presence of activated neutrophils or purified proteinase 3. Proc. Natl. Acad. Sci. USA96, 6261–6266.10.1073/pnas.96.11.6261Search in Google Scholar

Coligan, J., Dunn, B., Ploegh, H., Speicher, D., and Wingfield, P. (1997). Current Protocols in Protein Science (New York, USA: John Wiley & Sons), pp. 6.6.5–6.6.10.Search in Google Scholar

Desrochers, P.E., Mookhtiar, K., Van Wart, H.E., Hasty, K.A., and Weiss, S.J. (1992). Proteolytic inactivation of alpha 1-proteinase inhibitor and α1-antichymotrypsin by oxidatively activated human neutrophil metalloproteinases. J. Biol. Chem.267, 5005–5012.10.1016/S0021-9258(18)42931-6Search in Google Scholar

Dollery, C.M., Owen, C.A., Sukhova, G.K., Krettek, A., Shapiro, S.D., and Libby, P. (2003). Neutrophil elastase in human atherosclerotic plaques: production by macrophages. Circulation107, 2829–2836.10.1161/01.CIR.0000072792.65250.4ASearch in Google Scholar

Doring, G., Frank, F., Boudier, C., Herbert, S., Fleischer, B., and Bellon, G. (1995). Cleavage of lymphocyte surface antigens CD2, CD4, and CD8 by polymorphonuclear leukocyte elastase and cathepsin G in patients with cystic fibrosis. J. Immunol.154, 4842–4850.Search in Google Scholar

Eissner, G., Kohlhuber, F., Grell, M., Ueffing, M., Scheurich, P., Hieke, A., Multhoff, G., Bornkamm, G.W., and Holler, E. (1995). Critical involvement of transmembrane tumor necrosis factor α in endothelial programmed cell death mediated by ionizing radiation and bacterial endotoxin. Blood86, 4184–4193.10.1182/blood.V86.11.4184.bloodjournal86114184Search in Google Scholar

Eissner, G., Kolch, W., and Scheurich, P. (2004). Ligands working as receptors: reverse signaling by members of the TNF superfamily enhance the plasticity of the immune system. Cytokine Growth Factor Rev.15, 353–366.10.1016/j.cytogfr.2004.03.011Search in Google Scholar

Gasparini, C., Menegazzi, R., Patriarca, P., and Dri, P. (2003). Evidence that elastase is the TNF-R75 shedding enzyme in resting human polymorphonuclear leukocytes. FEBS Lett.553, 360–364.10.1016/S0014-5793(03)01046-9Search in Google Scholar

Ginzberg, H.H., Cherapanov, V., Dong, Q., Cantin, A., McCulloch, C.A., Shannon, P.T., and Downey, G.P. (2001). Neutrophil-mediated epithelial injury during transmigration: role of elastase. Am. J. Physiol. Gastrointest. Liver Physiol.281, G705–717.10.1152/ajpgi.2001.281.3.G705Search in Google Scholar

Goh, C.R. and Porter, A.G. (1991). Structural and functional domains in human tumour necrosis factors. Protein Eng.4, 385–389.10.1093/protein/4.4.385Search in Google Scholar

Grell, M., Douni, E., Wajant, H., Lohden, M., Clauss, M., Maxeiner, B., Georgopoulos, S., Lesslauer, W., Kollias, G., Pfizenmaier, K., et al. (1995). The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor. Cell83, 793–802.10.1016/0092-8674(95)90192-2Search in Google Scholar

Grell, M., Zimmermann, G., Gottfried, E., Chen, C.M., Grunwald, U., Huang, D.C., Wu Lee, Y.H., Durkop, H., Engelmann, H., Scheurich, P., Wajant, H., and Strasser, A. (1999). Induction of cell death by tumour necrosis factor (TNF) receptor 2, CD40 and CD30: a role for TNF-R1 activation by endogenous membrane-anchored TNF. EMBO J.18, 3034–3043.10.1093/emboj/18.11.3034Search in Google Scholar

Harigai, M., Hara, M., Nakazawa, S., Fukasawa, C., Ohta, S., Sugiura, T., Inoue, K., and Kashiwazaki, S. (1999). Ligation of CD40 induced tumor necrosis factor-α in rheumatoid arthritis: a novel mechanism of activation of synoviocytes. J. Rheumatol.26, 1035–1043.Search in Google Scholar

Hehlgans, T. and Mannel, D.N. (2002). The TNF-TNF receptor system. Biol. Chem.383, 1581–1585.Search in Google Scholar

Johnson, D. and Travis, J. (1979). The oxidative inactivation of human α-1-proteinase inhibitor. Further evidence for methionine at the reactive center. J. Biol. Chem.254, 4022–4026.Search in Google Scholar

Lazdins, J.K., Grell, M., Walker, M.R., Woods-Cook, K., Scheurich, P., and Pfizenmaier, K. (1997). Membrane tumor necrosis factor (TNF) induced cooperative signaling of TNFR60 and TNFR80 favors induction of cell death rather than virus production in HIV-infected T cells. J. Exp. Med.185, 81–90.10.1084/jem.185.1.81Search in Google Scholar

Le-Barillec, K., Si-Tahar, M., Balloy, V., and Chignard, M. (1999). Proteolysis of monocyte CD14 by human leukocyte elastase inhibits lipopolysaccharide-mediated cell activation. J. Clin. Invest.103, 1039–1046.10.1172/JCI5779Search in Google Scholar

Liou, T.G. and Campbell, E.J. (1996). Quantum proteolysis resulting from release of single granules by human neutrophils: a novel, nonoxidative mechanism of extracellular proteolytic activity. J. Immunol.157, 2624–2631.Search in Google Scholar

MacEwan, D.J. (2002). TNF receptor subtype signalling: differences and cellular consequences. Cell Signal.14, 477–492.10.1016/S0898-6568(01)00262-5Search in Google Scholar

Marra, L.E., Zhang, Z.X., Joe, B., Campbell, J., Levy, G.A., Penninger, J., and Zhang, L. (2004). IL-10 induces regulatory T cell apoptosis by up-regulation of the membrane form of TNF-α. J. Immunol.172, 1028–1035.10.4049/jimmunol.172.2.1028Search in Google Scholar

Matsudaira, P. (1987). Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J. Biol. Chem.262, 10035–10038.10.1016/S0021-9258(18)61070-1Search in Google Scholar

Mezyk-Kopec, R., Bzowska, M., Potempa, J., Bzowska, M., Jura, N., Sroka, A., Black, R.A., and Bereta, J. (2005). Inactivation of membrane TNFα by gingipains from Porphyromonas gingivalis. Infect. Immun.73, 1506–1514.10.1128/IAI.73.3.1506-1514.2005Search in Google Scholar

Mohammed, F.F., Smookler, D.S., Taylor, S.E., Fingleton, B., Kassiri, Z., Sanchez, O.H., English, J.L., Matrisian, L.M., Au, B., Yeh, W.C., and Khokha, R. (2004). Abnormal TNF activity in Timp3-/- mice leads to chronic hepatic inflammation and failure of liver regeneration. Nat. Genet.36, 969–977.10.1038/ng1413Search in Google Scholar

Momohara, S., Kashiwazaki, S., Inoue, K., Saito, S., and Nakagawa, T. (1997). Elastase from polymorphonuclear leukocyte in articular cartilage and synovial fluids of patients with rheumatoid arthritis. Clin. Rheumatol.16, 133–140.10.1007/BF02247841Search in Google Scholar

Morrison, H.M., Welgus, H.G., Stockley, R.A., Burnett, D., and Campbell, E.J. (1990). Inhibition of human leukocyte elastase bound to elastin: relative ineffectiveness and two mechanisms of inhibitory activity. Am. J. Respir. Cell Mol. Biol.2, 263–269.10.1165/ajrcmb/2.3.263Search in Google Scholar

Mueller, C., Corazza, N., Trachsel-Loseth, S., Eugster, H.P., Buhler-Jungo, M., Brunner, T., and Imboden, M.A. (1999). Noncleavable transmembrane mouse tumor necrosis factor-alpha (TNFalpha) mediates effects distinct from those of wild-type TNFα in vitro and in vivo. J. Biol. Chem.274, 38112–38118.10.1074/jbc.274.53.38112Search in Google Scholar

Nakamura, H., Yoshimura, K., McElvaney, N.G., and Crystal, R.G. (1992). Neutrophil elastase in respiratory epithelial lining fluid of individuals with cystic fibrosis induces interleukin-8 gene expression in a human bronchial epithelial cell line. J. Clin. Invest.89, 1478–1484.10.1172/JCI115738Search in Google Scholar

Nie, J. and Pei, D. (2004). Rapid inactivation of α-1-proteinase inhibitor by neutrophil specific leukolysin/membrane-type matrix metalloproteinase 6. Exp. Cell Res.296, 145–150.10.1016/j.yexcr.2004.02.008Search in Google Scholar

Omura, T., Yoshiyama, M., Kim, S., Matsumoto, R., Nakamura, Y., Izumi, Y., Ichijo, H., Sudo, T., Akioka, K., Iwao, H., Takeuchi, K., and Yoshikawa, J. (2004). Involvement of apoptosis signal-regulating kinase-1 on angiotensin II-induced monocyte chemoattractant protein-1 expression. Arterioscler. Thromb. Vasc. Biol.24, 270–275.10.1161/01.ATV.0000112930.40564.89Search in Google Scholar

Owen, C.A. and Campbell, E.J. (1998). Angiotensin II generation at the cell surface of activated neutrophils: novel cathepsin G-mediated catalytic activity that is resistant to inhibition. J. Immunol.160, 1436–1443.Search in Google Scholar

Owen, C.A., Campbell, M.A., Sannes, P.L., Boukedes, S.S., and Campbell, E.J. (1995). Cell surface-bound elastase and cathepsin G on human neutrophils: a novel, non-oxidative mechanism by which neutrophils focus and preserve catalytic activity of serine proteinases. J. Cell. Biol.131, 775–789.10.1083/jcb.131.3.775Search in Google Scholar

Owen, C.A., Campbell, M.A., Boukedes, S.S., and Campbell, E.J. (1997). Cytokines regulate membrane-bound leukocyte elastase on neutrophils: a novel mechanism for effector activity. Am. J. Physiol.272, L385–393.10.1152/ajplung.1997.272.3.L385Search in Google Scholar

Peters, J. and Baumgarten, H. (1992). Monoclonal Antibodies: A Practical Guide (New York, USA: Springer-Verlag).10.1007/978-3-642-74532-4Search in Google Scholar

Pikarsky, E., Porat, R.M., Stein, I., Abramovitch, R., Amit, S., Kasem, S., Gutkovich-Pyest, E., Urieli-Shoval, S., Galun, E., and Ben-Neriah, Y. (2004). NF-κB functions as a tumour promoter in inflammation-associated cancer. Nature431, 461–466.10.1038/nature02924Search in Google Scholar

Porteu, F., Brockhaus, M., Wallach, D., Engelmann, H., and Nathan, C.F. (1991). Human neutrophil elastase releases a ligand-binding fragment from the 75-kDa tumor necrosis factor (TNF) receptor. Comparison with the proteolytic activity responsible for shedding of TNF receptors from stimulated neutrophils. J. Biol. Chem.266, 18846–18853.Search in Google Scholar

Robache-Gallea, S., Morand, V., Bruneau, J.M., Schoot, B., Tagat, E., Realo, E., Chouaib, S., and Roman-Roman, S. (1995). In vitro processing of human tumor necrosis factor-α. J. Biol. Chem.270, 23688–23692.10.1074/jbc.270.40.23688Search in Google Scholar

Rocha-Pereira, P., Santos-Silva, A., Rebelo, I., Figueiredo, A., Quintanilha, A., and Teixeira, F. (2004). The inflammatory response in mild and in severe psoriasis. Br. J. Dermatol.150, 917–928.10.1111/j.1365-2133.2004.05984.xSearch in Google Scholar

Sadallah, S., Hess, C., Miot, S., Spertini, O., Lutz, H., and Schifferli, J.A. (1999). Elastase and metalloproteinase activities regulate soluble complement receptor 1 release. Eur. J. Immunol.29, 3754–3761.10.1002/(SICI)1521-4141(199911)29:11<3754::AID-IMMU3754>3.0.CO;2-5Search in Google Scholar

Schägger, H. and von Jagow, G. (1987). Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem.166, 368–379.10.1016/0003-2697(87)90587-2Search in Google Scholar

Schmid, E.F., Binder, K., Grell, M., Scheurich, P., and Pfizenmaier, K. (1995). Both tumor necrosis factor receptors, TNFR60 and TNFR80, are involved in signaling endothelial tissue factor expression by juxtacrine tumor necrosis factor α. Blood86, 1836–1841.10.1182/blood.V86.5.1836.bloodjournal8651836Search in Google Scholar

Shapiro, S.D. (2002). Neutrophil elastase: path clearer, pathogen killer, or just pathologic? Am. J. Respir. Cell Mol. Biol.26, 266–268.10.1165/ajrcmb.26.3.f233Search in Google Scholar

Stockley, R.A. (1999). Neutrophils and protease/antiprotease imbalance. Am. J. Respir. Crit. Care Med.160, S49–52.10.1164/ajrccm.160.supplement_1.13Search in Google Scholar

Sun, Z. and Yang, P. (2004). Role of imbalance between neutrophil elastase and α1-antitrypsin in cancer development and progression. Lancet Oncol.5, 182–190.10.1016/S1470-2045(04)01414-7Search in Google Scholar

Tosi, M.F., Zakem, H., and Berger, M. (1990). Neutrophil elastase cleaves C3bi on opsonized Pseudomonas as well as CR1 on neutrophils to create a functionally important opsonin receptor mismatch. J. Clin. Invest.86, 300–308.10.1172/JCI114699Search in Google Scholar

van Kessel, K.P., van Strijp, J.A., and Verhoef, J. (1991). Inactivation of recombinant human tumor necrosis factor α by proteolytic enzymes released from stimulated human neutrophils. J. Immunol.147, 3862–3868.Search in Google Scholar

Waetzig, G.H., Rosenstiel, P., Arlt, A., Till, A., Brautigam, K., Schafer, H., Rose-John, S., Seegert, D., and Schreiber, S. (2005). Soluble tumor necrosis factor (TNF) receptor-1 induces apoptosis via reverse TNF signaling and autocrine transforming growth factor β1. FASEB J.19, 91–93.10.1096/fj.04-2073fjeSearch in Google Scholar

Weiss, S.J. (1989). Tissue destruction by neutrophils. N. Engl. J. Med.320, 365–76.Search in Google Scholar

Weiss, T., Grell, M., Hessabi, B., Bourteele, S., Muller, G., Scheurich, P., and Wajant, H. (1997). Enhancement of TNF receptor p60-mediated cytotoxicity by TNF receptor p80: requirement of the TNF receptor-associated factor-2 binding site. J. Immunol.158, 2398–2404.Search in Google Scholar

Wright, G., Singh, I.S., Hasday, J.D., Farrance, I.K., Hall, G., Cross, A.S., and Rogers, T.B. (2002). Endotoxin stress-response in cardiomyocytes: NF-κB activation and tumor necrosis factor α expression. Am. J. Physiol. Heart Circ. Physiol.282, H872–879.10.1152/ajpheart.00256.2001Search in Google Scholar

Zhu, Y.K., Liu, X.D., Skold, C.M., Umino, T., Wang, H.J., Spurzem, J.R., Kohyama, T., Ertl, R.F., and Rennard, S.I. (2001). Synergistic neutrophil elastase-cytokine interaction degrades collagen in three-dimensional culture. Am. J. Physiol. Lung Cell. Mol. Physiol.281, L868–878.10.1152/ajplung.2001.281.4.L868Search in Google Scholar

Published Online: 2005-09-06
Published in Print: 2005-08-01

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