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. 1996 Sep;64(9):3491–3496. doi: 10.1128/iai.64.9.3491-3496.1996

Partial characterization of a cell proliferation-inhibiting protein produced by Helicobacter pylori.

U Knipp 1, S Birkholz 1, W Kaup 1, W Opferkuch 1
PMCID: PMC174253  PMID: 8751889

Abstract

Despite the induction of an immunological reaction, Helicobacter pylori-associated gastritis is a chronic disease, suggesting that this microbe can evade the host immune defense. Previous studies by our group showed that H. pylori suppresses the in vitro proliferative response of human mononuclear cells to mitogens and antigens. Here we demonstrate that the antiproliferative activity of H. pylori also affects the proliferation of various mammalian cell lines (U937, Jurkat, AGS, Kato-3, HEP-2, and P388D1). This effect is detectable in the first 16 h of incubation and maximal between 24 and 48 h. In addition, the presence of H. pylori significantly diminished the protein synthesis of cells in the first 6 h of incubation, comparable to the results with cycloheximide and diphtheria toxin. The urease enzyme, the cagA gene product, and the vacuolizing cytotoxin of H. pylori were excluded as causative agents of the antiproliferative effect by using isogenic knockout mutant strains. The inhibitory effect was not due to a lytic activity of this bacterium. The results reported here indicate that the responsible factor is a protein with an apparent native molecular mass of 100 +/- 10 kDa. Our work implicates the presence of a protein factor in H. pylori (termed PIP [for proliferation-inhibiting protein]) with antiproliferative activity for mammalian cells, including immunocompetent and epithelial cells. Thus, it is reasonable to presume that this property may contribute to the pathogenesis of H. pylori-induced diseases. It may be involved on the one hand in immune response evasion and on the other hand in the suppression of epithelial repair mechanisms.

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Selected References

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  1. Benz J., Hasbach H., Brenden M., Eidt S., Fätkenheuer G., Schrappe M., Geisel J., Goossens H., Mauff G. Humoral and cellular immunity in HIV positive and HIV negative Helicobacter pylori infected patients. Zentralbl Bakteriol. 1993 Sep;280(1-2):186–196. doi: 10.1016/s0934-8840(11)80955-2. [DOI] [PubMed] [Google Scholar]
  2. Blaser M. J. Helicobacter pylori and the pathogenesis of gastroduodenal inflammation. J Infect Dis. 1990 Apr;161(4):626–633. doi: 10.1093/infdis/161.4.626. [DOI] [PubMed] [Google Scholar]
  3. Blaser M. J. Helicobacter pylori: microbiology of a 'slow' bacterial infection. Trends Microbiol. 1993 Oct;1(7):255–260. doi: 10.1016/0966-842x(93)90047-u. [DOI] [PubMed] [Google Scholar]
  4. Blaser M. J. Hypotheses on the pathogenesis and natural history of Helicobacter pylori-induced inflammation. Gastroenterology. 1992 Feb;102(2):720–727. doi: 10.1016/0016-5085(92)90126-j. [DOI] [PubMed] [Google Scholar]
  5. Buck G. E. Campylobacter pylori and gastroduodenal disease. Clin Microbiol Rev. 1990 Jan;3(1):1–12. doi: 10.1128/cmr.3.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chang M. P., Bramhall J., Graves S., Bonavida B., Wisnieski B. J. Internucleosomal DNA cleavage precedes diphtheria toxin-induced cytolysis. Evidence that cell lysis is not a simple consequence of translation inhibition. J Biol Chem. 1989 Sep 15;264(26):15261–15267. [PubMed] [Google Scholar]
  7. Chow K. W., Bank S., Ahn J., Roberts J., Blumstein M., Kranz V. Helicobacter pylori infection does not increase gastric antrum mucosal cell proliferation. Am J Gastroenterol. 1995 Jan;90(1):64–66. [PubMed] [Google Scholar]
  8. Covacci A., Censini S., Bugnoli M., Petracca R., Burroni D., Macchia G., Massone A., Papini E., Xiang Z., Figura N. Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori associated with cytotoxicity and duodenal ulcer. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5791–5795. doi: 10.1073/pnas.90.12.5791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cover T. L., Blaser M. J. Purification and characterization of the vacuolating toxin from Helicobacter pylori. J Biol Chem. 1992 May 25;267(15):10570–10575. [PubMed] [Google Scholar]
  10. Cover T. L., Dooley C. P., Blaser M. J. Characterization of and human serologic response to proteins in Helicobacter pylori broth culture supernatants with vacuolizing cytotoxin activity. Infect Immun. 1990 Mar;58(3):603–610. doi: 10.1128/iai.58.3.603-610.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cover T. L., Puryear W., Perez-Perez G. I., Blaser M. J. Effect of urease on HeLa cell vacuolation induced by Helicobacter pylori cytotoxin. Infect Immun. 1991 Apr;59(4):1264–1270. doi: 10.1128/iai.59.4.1264-1270.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dixon M. F., Wyatt J. I., Burke D. A., Rathbone B. J. Lymphocytic gastritis--relationship to Campylobacter pylori infection. J Pathol. 1988 Feb;154(2):125–132. doi: 10.1002/path.1711540204. [DOI] [PubMed] [Google Scholar]
  13. Ferrero R. L., Cussac V., Courcoux P., Labigne A. Construction of isogenic urease-negative mutants of Helicobacter pylori by allelic exchange. J Bacteriol. 1992 Jul;174(13):4212–4217. doi: 10.1128/jb.174.13.4212-4217.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Geis G., Leying H., Suerbaum S., Mai U., Opferkuch W. Ultrastructure and chemical analysis of Campylobacter pylori flagella. J Clin Microbiol. 1989 Mar;27(3):436–441. doi: 10.1128/jcm.27.3.436-441.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hu L. T., Mobley H. L. Purification and N-terminal analysis of urease from Helicobacter pylori. Infect Immun. 1990 Apr;58(4):992–998. doi: 10.1128/iai.58.4.992-998.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hussell T., Isaacson P. G., Crabtree J. E., Spencer J. The response of cells from low-grade B-cell gastric lymphomas of mucosa-associated lymphoid tissue to Helicobacter pylori. Lancet. 1993 Sep 4;342(8871):571–574. doi: 10.1016/0140-6736(93)91408-e. [DOI] [PubMed] [Google Scholar]
  17. Knipp U., Birkholz S., Kaup W., Mahnke K., Opferkuch W. Suppression of human mononuclear cell response by Helicobacter pylori: effects on isolated monocytes and lymphocytes. FEMS Immunol Med Microbiol. 1994 Feb;8(2):157–166. doi: 10.1111/j.1574-695X.1994.tb00438.x. [DOI] [PubMed] [Google Scholar]
  18. Knipp U., Birkholz S., Kaup W., Opferkuch W. Immune suppressive effects of Helicobacter pylori on human peripheral blood mononuclear cells. Med Microbiol Immunol. 1993 May;182(2):63–76. doi: 10.1007/BF00189374. [DOI] [PubMed] [Google Scholar]
  19. Lee A., Fox J., Hazell S. Pathogenicity of Helicobacter pylori: a perspective. Infect Immun. 1993 May;61(5):1601–1610. doi: 10.1128/iai.61.5.1601-1610.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Leunk R. D., Johnson P. T., David B. C., Kraft W. G., Morgan D. R. Cytotoxic activity in broth-culture filtrates of Campylobacter pylori. J Med Microbiol. 1988 Jun;26(2):93–99. doi: 10.1099/00222615-26-2-93. [DOI] [PubMed] [Google Scholar]
  21. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  22. Lynch D. A., Mapstone N. P., Clarke A. M., Jackson P., Dixon M. F., Quirke P., Axon A. T. Cell proliferation in the gastric corpus in Helicobacter pylori associated gastritis and after gastric resection. Gut. 1995 Mar;36(3):351–353. doi: 10.1136/gut.36.3.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mobley H. L., Cortesia M. J., Rosenthal L. E., Jones B. D. Characterization of urease from Campylobacter pylori. J Clin Microbiol. 1988 May;26(5):831–836. doi: 10.1128/jcm.26.5.831-836.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. doi: 10.1016/0022-1759(83)90303-4. [DOI] [PubMed] [Google Scholar]
  25. Mégraud F., Neman-Simha V., Brügmann D. Further evidence of the toxic effect of ammonia produced by Helicobacter pylori urease on human epithelial cells. Infect Immun. 1992 May;60(5):1858–1863. doi: 10.1128/iai.60.5.1858-1863.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Parsonnet J., Friedman G. D., Vandersteen D. P., Chang Y., Vogelman J. H., Orentreich N., Sibley R. K. Helicobacter pylori infection and the risk of gastric carcinoma. N Engl J Med. 1991 Oct 17;325(16):1127–1131. doi: 10.1056/NEJM199110173251603. [DOI] [PubMed] [Google Scholar]
  27. Perentesis J. P., Miller S. P., Bodley J. W. Protein toxin inhibitors of protein synthesis. Biofactors. 1992 Jan;3(3):173–184. [PubMed] [Google Scholar]
  28. Phadnis S. H., Parlow M. H., Levy M., Ilver D., Caulkins C. M., Connors J. B., Dunn B. E. Surface localization of Helicobacter pylori urease and a heat shock protein homolog requires bacterial autolysis. Infect Immun. 1996 Mar;64(3):905–912. doi: 10.1128/iai.64.3.905-912.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rathbone B. J., Wyatt J. I., Worsley B. W., Shires S. E., Trejdosiewicz L. K., Heatley R. V., Losowsky M. S. Systemic and local antibody responses to gastric Campylobacter pyloridis in non-ulcer dyspepsia. Gut. 1986 Jun;27(6):642–647. doi: 10.1136/gut.27.6.642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schmitt W., Haas R. Genetic analysis of the Helicobacter pylori vacuolating cytotoxin: structural similarities with the IgA protease type of exported protein. Mol Microbiol. 1994 Apr;12(2):307–319. doi: 10.1111/j.1365-2958.1994.tb01019.x. [DOI] [PubMed] [Google Scholar]
  31. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  32. Smoot D. T., Mobley H. L., Chippendale G. R., Lewison J. F., Resau J. H. Helicobacter pylori urease activity is toxic to human gastric epithelial cells. Infect Immun. 1990 Jun;58(6):1992–1994. doi: 10.1128/iai.58.6.1992-1994.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stolte M., Eidt S. Lymphoid follicles in antral mucosa: immune response to Campylobacter pylori? J Clin Pathol. 1989 Dec;42(12):1269–1271. doi: 10.1136/jcp.42.12.1269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Taylor D. N., Blaser M. J. The epidemiology of Helicobacter pylori infection. Epidemiol Rev. 1991;13:42–59. doi: 10.1093/oxfordjournals.epirev.a036078. [DOI] [PubMed] [Google Scholar]
  35. Telford J. L., Covacci A., Ghiara P., Montecucco C., Rappuoli R. Unravelling the pathogenic role of Helicobacter pylori in peptic ulcer: potential new therapies and vaccines. Trends Biotechnol. 1994 Oct;12(10):420–426. doi: 10.1016/0167-7799(94)90031-0. [DOI] [PubMed] [Google Scholar]
  36. Wotherspoon A. C., Doglioni C., Diss T. C., Pan L., Moschini A., de Boni M., Isaacson P. G. Regression of primary low-grade B-cell gastric lymphoma of mucosa-associated lymphoid tissue type after eradication of Helicobacter pylori. Lancet. 1993 Sep 4;342(8871):575–577. doi: 10.1016/0140-6736(93)91409-f. [DOI] [PubMed] [Google Scholar]
  37. Wyatt J. I., Rathbone B. J., Heatley R. V. Local immune response to gastric Campylobacter in non-ulcer dyspepsia. J Clin Pathol. 1986 Aug;39(8):863–870. doi: 10.1136/jcp.39.8.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wyatt J. I., Rathbone B. J. Immune response of the gastric mucosa to Campylobacter pylori. Scand J Gastroenterol Suppl. 1988;142:44–49. [PubMed] [Google Scholar]
  39. Zhu L., Anasetti C. Cell cycle control of apoptosis in human leukemic T cells. J Immunol. 1995 Jan 1;154(1):192–200. [PubMed] [Google Scholar]

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