Helicobacter pylori-induced downregulation of the secretory leukocyte protease inhibitor (SLPI) in gastric epithelial cell lines and its functional relevance for H. pylori-mediated diseases
-
Thomas Wex
, Gerhard Treiber , Marino Venerito , Andreas Leodolter , Ulrich Peitz , Doerthe Kuester , Istvan Hritz , Sabine Krueger , Albert Roessner and Peter Malfertheiner
Abstract
The secretory leukocyte protease inhibitor (SLPI) exerts antiproteolytic activity towards serine proteases, as well as anti-microbial and anti-inflammatory effects. To investigate its role in H. pylori-mediated diseases, SLPI expression was analyzed by RT-PCR, ELISA and immunohistochemistry in clinical samples and gastric tumor cell lines. Determination of the mucosal SLPI levels in 126 patients confirmed the previously reported downregulation of SLPI in H. pylori-infected patients. The lower SLPI levels in antral biopsies of H. pylori-positive subjects were associated with a 30-fold increase (p<0.01) in neutrophil elastase activity, and a significant negative correlation was demonstrated for both parameters (R=-0.63, p=0.0002). Eradication of the bacterium in a long-term study (5–7 years) led to a recovery of mucosal SLPI expression. In vitro experiments using four gastric tumor cell lines (AGS, MKN-28, MKN-45, NCI-N87) generally confirmed the clinical findings. While the co-incubation of these cell lines with H. pylori resulted in lower or unchanged SLPI protein levels, the corresponding SLPI mRNA amounts were upregulated by up to five-fold (p=0.006) in all cell lines. Taken together, these results indicate that the reduction in antral SLPI levels in H. pylori-infected subjects has a functional relevance for gastric mucosa and the H. pylori-induced decrease in SLPI is primarily regulated at the posttranslational level.
References
Abe, T., Kobayashi, N., Yoshimura, K., Trapnell, B.C., Kim, H., Hubbard, R.C., Brewer, M.T., Thompson, R.C., and Crystal R.G. (1991). Expression of the secretory leukoprotease inhibitor gene in epithelial cells. J. Clin. Invest.87, 2207–2215.10.1172/JCI115255Search in Google Scholar
Alkemade, J.A., Molhuizen, H.O., Ponec, M., Kempenaar, J.A., Zeeuwen, P.L., de Jongh, G.J., van Vlijmen-Willems, I.M., van Erp, P.E., Van de Kerkhof, P.C., and Schalkwijk, J. (1994). SKALP/elafin is an inducible proteinase inhibitor in human epidermal keratinocytes. J. Cell Sci.107, 2335–2342.10.1242/jcs.107.8.2335Search in Google Scholar
Ashcroft, G.S., Lei, K., Jin, W., Longenecker, G., Kulkarni, A.B., and Greenwell-Wild, T. (2000). Secretory leukocyte protease inhibitor mediates non-redundant functions necessary for normal wound healing. Nat. Med.6, 1147–1153.10.1038/80489Search in Google Scholar
Bergenfeldt, M., Nystrom, M., Bohe, M., Lindstrom, C., Polling, A., and Ohlsson, K. (1996). Localization of immunoreactive secretory leukocyte protease inhibitor (SLPI) in intestinal mucosa. J. Gastroenterol.31, 18–23.10.1007/BF01211182Search in Google Scholar
Boudier, C., Cadene, M., and Bieth, J.G. (1999). Inhibition of neutrophil cathepsin G by oxidized mucus proteinase inhibitor. Effect of heparin. Biochemistry38, 8451–8457.Search in Google Scholar
Brandt, S., Kwok, T., Hartig, R., Konig, W., and Backert, S. (2005). NF-κB activation and potentiation of proinflammatory responses by the Helicobacter pylori CagA protein. Proc. Natl. Acad. Sci. USA102, 9300–9305.10.1073/pnas.0409873102Search in Google Scholar
Buhling, F., Peitz, U., Kruger, S., Kuster, D., Vieth, M., Gebert, I., Roessner, A., Weber, E., Malfertheiner, P., and Wex, T. (2004). Cathepsins K, L, B, X and W are differentially expressed in normal and chronically inflamed gastric mucosa. Biol. Chem.385, 439–445.10.1515/BC.2004.051Search in Google Scholar
Chattopadhyay, A., Gray, L.R., Patton, L.L., Caplan, D.J., Slade, G.D., Tien, H.C., and Shugars, D.C. (2004). Salivary secretory leukocyte protease inhibitor and oral candidiasis in human immunodeficiency virus type 1-infected persons. Infect. Immun.72, 1956–1963.10.1128/IAI.72.4.1956-1963.2004Search in Google Scholar
Crabtree, J.E. (1996). Immune and inflammatory responses to Helicobacter pylori infection. Scand. J. Gastroenterol. (Suppl.) 215, 3–10.10.3109/00365529609094526Search in Google Scholar
Dixon, M.F., Genta, R.M., Yardley, J.H., and Correa, P. (1996). Classification and grading of gastritis. The updated Sydney System. International Workshop on the Histopathology of Gastritis, Houston 1994. Am. J. Surg. Pathol.20, 1161–1181.10.1097/00000478-199610000-00001Search in Google Scholar
Dohchin, A., Suzuki, J.I., Seki, H., Masutani, M., Shiroto, H., and Kawakami Y. (2000). Immunostained cathepsins B and L correlate with depth of invasion and different metastatic pathways in early stage gastric carcinoma. Cancer89, 482–487.10.1002/1097-0142(20000801)89:3<482::AID-CNCR2>3.0.CO;2-5Search in Google Scholar
Duits, L.A., Tjabringa, G.S., Aarts, N.J., van't Wout, J.W., Hiemstra, P.S., Nibbering, P.H., and van Dissel, J.T. (2003). Plasma secretory leukocyte protease inhibitor in febrile patients. Clin. Microbiol. Infect.9, 605–613.10.1046/j.1469-0691.2003.00582.xSearch in Google Scholar
Figueiredo, C., Machado, J.C., and Yamaoka, Y. (2005). Pathogenesis of Helicobacter pylori infection. Helicobacter10 (Suppl. 1), 14–20.10.1111/j.1523-5378.2005.00339.xSearch in Google Scholar
Fink, E., Nettelbeck, R., and Fritz, H. (1986). Inhibition of mast cell chymase by eglin c and antileukoprotease (HUSI-I). Indications for potential biological functions of these inhibitors. Biol. Chem. Hoppe-Seyler367, 567–571.Search in Google Scholar
Fritz, H. (1988). Human mucus proteinase inhibitor (human MPI). Human seminal inhibitor I (HUSI-I), antileukoprotease (ALP), secretory leukocyte protease inhibitor (SLPI). Biol. Chem. Hoppe-Seyler369, 79–82.Search in Google Scholar
Gils, A. and Declerck, P.J. (1998). Structure-function relationships in serpins: current concepts and controversies. Thromb. Haemost.80, 531–541.Search in Google Scholar
Grobmyer, S.R., Barie, P.S., Nathan, C.F., Fuortes, M., Lin, E., Lowry, S.F., Wright, C.D., Weyant, M.J., Hydo, L., Reeves, F., Shiloh, M.U., and Ding, A. (2000). Secretory leukocyte protease inhibitor, an inhibitor of neutrophil activation, is elevated in serum in human sepsis and experimental endotoxemia. Crit. Care Med.28, 1276–1282.10.1097/00003246-200005000-00003Search in Google Scholar
Heinzel, R., Appelhans, H., Gassen, G., Seemuller, U., Machleidt, W., Fritz, H., and Steffens, G. (1986). Molecular cloning and expression of cDNA for human antileukoprotease from cervix uterus. Eur. J. Biochem.160, 61–67.10.1111/j.1432-1033.1986.tb09940.xSearch in Google Scholar
Henriksen, P.A., Hitt, M., Xing, Z., Wang, J., Haslett, C., Riemersma, R.A., Webb, D.J., Kotelevtsev, Y.V., and Sallenave, J.M. (2004). Adenoviral gene delivery of elafin and secretory leukocyte protease inhibitor attenuates NF-κB-dependent inflammatory responses of human endothelial cells and macrophages to atherogenic stimuli. J. Immunol.172, 4535–4544.10.4049/jimmunol.172.7.4535Search in Google Scholar
Hiemstra, P.S., Maassen, R.J., Stolk, J., Heinzel-Wieland, R., Steffens, G.J., and Dijkman, J.H. (1996). Antibacterial activity of antileukoprotease. Infect. Immun.64, 4520–4524.10.1128/iai.64.11.4520-4524.1996Search in Google Scholar
Hritz, I., Kuester, D., Vieth, M., Herszenyi, L., Stolte, M., Roessner, A., Tulassay, Z., Wex, T., and Malfertheiner, P. (2006). Secretory leukocyte protease inhibitor expression in various types of gastritis: a specific role of Helicobacter pylori infection. Eur. J. Gastroenterol. Hepatol.18, 277–282.10.1097/00042737-200603000-00008Search in Google Scholar
Itoh, T., Wakatsuki, Y., Yoshida, M., Usui, T., Matsunaga, Y., Kaneko, S., Chiba, T., and Kita, T. (1999). The vast majority of gastric T cells are polarized to produce T helper 1 type cytokines upon antigenic stimulation despite the absence of Helicobacter pylori infection. J. Gastroenterol.34, 560–570.10.1007/s005350050373Search in Google Scholar
Jones, B.D. (2005). Salmonella invasion gene regulation: a story of environmental awareness. J. Microbiol.43, 110–117.Search in Google Scholar
Koyama, S. (2004). Significance of cell-surface expression of matrix metalloproteinases and their inhibitors on gastric epithelium and infiltrating mucosal lymphocytes in progression of Helicobacter pylori-associated gastritis. Scand. J. Gastroenterol.39, 1046–1053.10.1080/00365520410003245Search in Google Scholar
Lentsch, A.B., Jordan, J.A., Czermak, B.J., Diehl, K.M., Younkin, E.M., Sarma, V., and Ward, P.A. (1999). Inhibition of NF-κB activation and augmentation of IκBβ by secretory leukocyte protease inhibitor during lung inflammation. Am. J. Pathol.154, 239–247.10.1016/S0002-9440(10)65270-4Search in Google Scholar
Lindholm, C., Quiding-Jarbrink, M., Lonroth, H., and Svennerholm, A.M. (2001). Induction of chemokine and cytokine responses by Helicobacter pylori in human stomach explants. Scand. J. Gastroenterol.36, 1022–1029.Search in Google Scholar
Naumann, M. (2001). Host cell signaling in Helicobacter pylori infection. Int. J. Med. Microbiol.291, 299–305.10.1078/1438-4221-00133Search in Google Scholar
Parsonnet, J., Friedman, G.D., Vandersteen, D.P., Chang, Y., Vogelman, J.H., Orentreich, N., and Sibley, R.K. (1991). Helicobacter pylori infection and the risk of gastric carcinoma. N. Engl. J. Med.325, 1127–1131.10.1056/NEJM199110173251603Search in Google Scholar
Potempa, J., Korzus, E., and Travis, J. (1994). The serpin superfamily of proteinase inhibitors: structure, function, and regulation. J. Biol. Chem.269, 15957–15960.10.1016/S0021-9258(17)33954-6Search in Google Scholar
Quiding-Jarbrink, M., Lundin, B.S., Lonroth, H., and Svennerholm, A.M. (2001). CD4+ and CD8+ T cell responses in Helicobacter pylori-infected individuals. Clin. Exp. Immunol.123, 81–87.10.1046/j.1365-2249.2001.01427.xSearch in Google Scholar
Sallenave, J.M. (2000). The role of secretory leukocyte proteinase inhibitor and elafin (elastase-specific inhibitor/skin-derived antileukoprotease) as alarm antiproteinases in inflammatory lung disease. Respir. Res.1, 87–92.10.1186/rr18Search in Google Scholar
Singh, P.K., Tack, B.F., McCray, P.B. Jr., and Welsh, M.J. (2000) Synergistic and additive killing by antimicrobial factors found in human airway surface liquid. Am. J. Physiol. Lung Cell. Mol. Physiol.279, 799–805.10.1152/ajplung.2000.279.5.L799Search in Google Scholar
Si-Tahar, M., Merlin, D., Sitaraman, S., and Madara, J.L. (2000). Constitutive and regulated secretion of secretory leukocyte proteinase inhibitor by human intestinal epithelial cells. Gastroenterology118, 1061–1071.10.1016/S0016-5085(00)70359-3Search in Google Scholar
Song, X., Zeng, L., Jin, W., Thompson, J., Mizel, D.E., Lei, K., Billinghurst, R.C., Poole, A.R., and Wahl, S.M. (1999). Secretory leukocyte protease inhibitor suppresses the inflammation and joint damage of bacterial cell wall-induced arthritis. J. Exp. Med.190, 535–542.10.1084/jem.190.4.535Search in Google Scholar PubMed PubMed Central
Suerbaum, S. and Michetti, P. (2002). Helicobacter pylori infection. N. Engl. J. Med.347, 1175–1186.10.1056/NEJMra020542Search in Google Scholar PubMed
Taggart, C.C., Lowe, G.J., Greene, C.M., Mulgrew, A.T., O'Neill, S.J., Levine, R.L., and McElvaney, N.G. (2000). Cathepsin B, L, and S cleave and inactivate secretory leucoprotease inhibitor. J. Biol. Chem.276, 33345–33352.Search in Google Scholar
Taggart, C.C., Greene, C.M., McElvaney, N.G., and O'Neill, S. (2002). Secretory leucoprotease inhibitor prevents LPS-induced IκBβ degradation without affecting phosphorylation or ubiquitination. J. Biol. Chem.277, 33648–33653.10.1074/jbc.M203710200Search in Google Scholar PubMed
Tam, E.M., Morrison, C.J., Wu, Y.I., Stack, M.S., and Overall, C.M. (2004). Membrane protease proteomics: isotope-coded affinity tag MS identification of undescribed MT1-matrix metalloproteinase substrates. Proc. Natl. Acad. Sci. USA101, 6917–6922.10.1073/pnas.0305862101Search in Google Scholar PubMed PubMed Central
Thompson, R.C. and Ohlsson, K. (1986). Isolation, properties, and complete amino acid sequence of human secretory leukocyte protease inhibitor, a potent inhibitor of leukocyte elastase. Proc. Natl. Acad. Sci. USA83, 6692–6696.10.1073/pnas.83.18.6692Search in Google Scholar PubMed PubMed Central
Toh, B.H., van Driel, I.R., and Gleeson, P.A. (1997). Pernicious anemia. N. Engl. J. Med.337, 1441–1448.10.1056/NEJM199711133372007Search in Google Scholar PubMed
Uemura, N., Okamoto, S., Yamamoto, S., Matsumura, N., Yamaguchi, S., Yamakido, M., Taniyama, K., Sasaki, N., and Schlemper, R.J. (2001). Helicobacter pylori infection and the development of gastric cancer. N. Engl. J. Med.345, 784–789.10.1056/NEJMoa001999Search in Google Scholar PubMed
Wang, X., Li, X., Xu, L., Zhan, Y., Yaish-Ohad, S., Erhardt, J.A., Barone, F.C., and Feuerstein, G.Z. (2003). Up-regulation of secretory leukocyte protease inhibitor (SLPI) in the brain after ischemic stroke: adenoviral expression of SLPI protects brain from ischemic injury. Mol. Pharmacol.64, 833–840.10.1124/mol.64.4.833Search in Google Scholar PubMed
Wex, T., Treiber, G., Lendeckel, U., and Malfertheiner, P. (2003). A two step method for the extraction of high-quality RNA from endoscopic biopsies. Clin. Chem. Lab. Med.41, 1033–1037.10.1515/CCLM.2003.159Search in Google Scholar PubMed
Wex, T., Treiber, G., Nilius, M., Vieth, M., Roessner, A., and Malfertheiner, P. (2004a). Helicobacter pylori-mediated gastritis induces local downregulation of secretory leukocyte protease inhibitor in the antrum. Infect. Immun.72, 2383–2385.10.1128/IAI.72.4.2383-2385.2004Search in Google Scholar PubMed PubMed Central
Wex, T., Sokic-Milutinovic, A., Todorovic, V., Bjelovic, M., Milosavljevic, T., Pesko, P., and Malfertheiner, P. (2004b). Down-regulation of secretory leukocyte protease inhibitor expression in gastric mucosa is a general phenomenon in Helicobacter pylori-related gastroduodenal diseases. Dig. Dis.22, 390–395.10.1159/000083604Search in Google Scholar PubMed
Wex, T., Zack, M., and Malfertheiner, P. (2006a). Proteases in Helicobacter pylori-mediated diseases. In: Proteases in Biology and Disease, Volume 5: Proteases in Gastrointestinal Tissues, U. Lendeckel and N.M. Hooper, eds. (New York, USA: Springer Science/Business Media Inc.), pp. 61–87.10.1007/1-4020-4483-6_3Search in Google Scholar
Wex, T., Ye, S., Treiber, G., Vieth, M., Roessner, A., and Malfertheiner, P. (2006b). H. pylori infection, but not low-dose aspirin, results in a local reduction of the secretory leukocyte protease inhibitor (SLPI) in gastroduodenal mucosa. Helicobacter11, 31–38.10.1111/j.0083-8703.2006.00376.xSearch in Google Scholar PubMed
Wiedow, O. and Meyer-Hoffert, U. (2005). Neutrophil serine proteases: potential key regulators of cell signaling during inflammation. J. Intern. Med.257, 319–328.10.1111/j.1365-2796.2005.01476.xSearch in Google Scholar
Yang, J., Zhu, J., Sun, D., and Ding, A. (2005). Suppression of macrophage responses to bacterial lipopolysaccharide (LPS) by secretory leukocyte protease inhibitor (SLPI) is independent of its anti-protease function. Biochim. Biophys. Acta1745, 310–317.10.1016/j.bbamcr.2005.07.006Search in Google Scholar
Zhang, D., Simmen, R.C., Michel, F.J., Zhao, G., Vale-Cruz, D., and Simmen, F.A. (2002). Secretory leukocyte protease inhibitor mediates proliferation of human endometrial epithelial cells by positive and negative regulation of growth-associated genes. J. Biol. Chem.277, 29999–30009.10.1074/jbc.M203503200Search in Google Scholar
Zarrilli, R., Ricci, V., and Romano, M. (1999). Molecular response of gastric epithelial cells to Helicobacter pylori-induced cell damage. Cell Microbiol.1, 93–99.10.1046/j.1462-5822.1999.00018.xSearch in Google Scholar
Zhu, J., Nathan, C., Jin, W., Sim, D., Ashcroft, G.S., Wahl, S.M., Lacomis, L., Erdjument-Bromage, H., Tempst, P., Wright, C.D., and Ding, A. (2002). Conversion of proepithelin to epithelia: roles of SLPI and elastase in host defense and wound repair. Cell111, 867–878.10.1016/S0092-8674(02)01141-8Search in Google Scholar
©2006 by Walter de Gruyter Berlin New York
Articles in the same Issue
- 4th General Meeting of the International Proteolysis Society/International Conference on Protease Inhibitors
- Extracellular granzymes: current perspectives
- Impact of the N-terminal amino acid on targeted protein degradation
- Structural aspects of recently discovered viral deubiquitinating activities
- Cysteine cathepsins and caspases in silicosis
- The proprotein convertases and their implication in sterol and/or lipid metabolism
- PREPL: a putative novel oligopeptidase propelled into the limelight
- Human cathepsin L rescues the neurodegeneration and lethality in cathepsin B/L double-deficient mice
- Helicobacter pylori-induced downregulation of the secretory leukocyte protease inhibitor (SLPI) in gastric epithelial cell lines and its functional relevance for H. pylori-mediated diseases
- Increased local levels of granulocyte colony-stimulating factor are associated with the beneficial effect of pre-elafin (SKALP/trappin-2/WAP3) in experimental emphysema
- Interaction of a novel form of Pseudomonas aeruginosa alkaline protease (aeruginolysin) with interleukin-6 and interleukin-8
- Analysis of aldosterone-induced differential receptor-independent protein patterns using 2D-electrophoresis and mass spectrometry
- Modeling the 3D structure of wheat subtilisin/chymotrypsin inhibitor (WSCI). Probing the reactive site with two susceptible proteinases by time-course analysis and molecular dynamics simulations
- A stable analogue of glucose-dependent insulinotropic polypeptide, GIP(LysPAL16), enhances functional differentiation of mouse embryonic stem cells into cells expressing islet-specific genes and hormones
- Transcription factor FOXM1c is repressed by RB and activated by cyclin D1/Cdk4
- Despite its strong transactivation domain, transcription factor FOXM1c is kept almost inactive by two different inhibitory domains
- Inhibition of calcineurin by infusion of CsA causes hyperphosphorylation of tau and is accompanied by abnormal behavior in mice
- Isolation and properties of extracellular proteinases of Penicillium marneffei
- Isolation and comparative characterization of Ki-67 equivalent antibodies from the HuCAL® phage display library
Articles in the same Issue
- 4th General Meeting of the International Proteolysis Society/International Conference on Protease Inhibitors
- Extracellular granzymes: current perspectives
- Impact of the N-terminal amino acid on targeted protein degradation
- Structural aspects of recently discovered viral deubiquitinating activities
- Cysteine cathepsins and caspases in silicosis
- The proprotein convertases and their implication in sterol and/or lipid metabolism
- PREPL: a putative novel oligopeptidase propelled into the limelight
- Human cathepsin L rescues the neurodegeneration and lethality in cathepsin B/L double-deficient mice
- Helicobacter pylori-induced downregulation of the secretory leukocyte protease inhibitor (SLPI) in gastric epithelial cell lines and its functional relevance for H. pylori-mediated diseases
- Increased local levels of granulocyte colony-stimulating factor are associated with the beneficial effect of pre-elafin (SKALP/trappin-2/WAP3) in experimental emphysema
- Interaction of a novel form of Pseudomonas aeruginosa alkaline protease (aeruginolysin) with interleukin-6 and interleukin-8
- Analysis of aldosterone-induced differential receptor-independent protein patterns using 2D-electrophoresis and mass spectrometry
- Modeling the 3D structure of wheat subtilisin/chymotrypsin inhibitor (WSCI). Probing the reactive site with two susceptible proteinases by time-course analysis and molecular dynamics simulations
- A stable analogue of glucose-dependent insulinotropic polypeptide, GIP(LysPAL16), enhances functional differentiation of mouse embryonic stem cells into cells expressing islet-specific genes and hormones
- Transcription factor FOXM1c is repressed by RB and activated by cyclin D1/Cdk4
- Despite its strong transactivation domain, transcription factor FOXM1c is kept almost inactive by two different inhibitory domains
- Inhibition of calcineurin by infusion of CsA causes hyperphosphorylation of tau and is accompanied by abnormal behavior in mice
- Isolation and properties of extracellular proteinases of Penicillium marneffei
- Isolation and comparative characterization of Ki-67 equivalent antibodies from the HuCAL® phage display library