Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Nov;62(11):4981–4989. doi: 10.1128/iai.62.11.4981-4989.1994

Recombinant antigens prepared from the urease subunits of Helicobacter spp.: evidence of protection in a mouse model of gastric infection.

R L Ferrero 1, J M Thiberge 1, M Huerre 1, A Labigne 1
PMCID: PMC303216  PMID: 7927778

Abstract

Urease is an important virulence factor for gastric Helicobacter spp. To elucidate the efficacy of individual urease subunits to act as mucosal immunogens, the genes encoding the respective urease subunits (UreA and UreB) of Helicobacter pylori and Helicobacter felis were cloned in an expression vector (pMAL) and expressed in Escherichia coli cells as translational fusion proteins. The recombinant UreA and UreB proteins were purified by affinity and anion-exchange chromatography techniques and had predicted molecular masses of approximately 68 and 103 kDa, respectively. Western blotting (immunoblotting) studies indicated that the urease components of the fusion proteins were strongly immunogenic and were specifically recognized by polyclonal rabbit anti-Helicobacter sp. sera. The fusion proteins (50 micrograms) were used, in combination with a mucosal adjuvant (cholera toxin), to orogastrically immunize mice against H. felis infection. Gastric tissues from H. felis-challenged mice were assessed by the biopsy urease test and by histology. In mice immunized with recombinant H. felis UreB, 60% of animals (n = 7) were histologically negative for H. felis bacteria after challenge at 17 weeks. This compared with 25% (n = 8) for mice immunized with the heterologous H. pylori UreB antigen. Neither the homologous nor the heterologous UreA subunit elicited protective responses against H. felis infection in mice. The study demonstrated that a recombinant subunit antigen could induce an immunoprotective response against gastric Helicobacter infection.

Full text

PDF
4981

Images in this article

4985Image
on p.4985
4985Image
on p.4985
4985Image
on p.4985
4986Image
on p.4986
4987Image
on p.4987
4987Image
on p.4987

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Booth L., Holdstock G., MacBride H., Hawtin P., Gibson J. R., Ireland A., Bamforth J., DuBoulay C. E., Lloyd R. S., Pearson A. D. Clinical importance of Campylobacter pyloridis and associated serum IgG and IgA antibody responses in patients undergoing upper gastrointestinal endoscopy. J Clin Pathol. 1986 Feb;39(2):215–219. doi: 10.1136/jcp.39.2.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen M., Lee A., Hazell S., Hu P., Li Y. Immunisation against gastric infection with Helicobacter species: first step in the prophylaxis of gastric cancer? Zentralbl Bakteriol. 1993 Sep;280(1-2):155–165. doi: 10.1016/s0934-8840(11)80952-7. [DOI] [PubMed] [Google Scholar]
  3. Chen M., Lee A., Hazell S. Immunisation against gastric helicobacter infection in a mouse/Helicobacter felis model. Lancet. 1992 May 2;339(8801):1120–1121. doi: 10.1016/0140-6736(92)90720-n. [DOI] [PubMed] [Google Scholar]
  4. Correa P. A human model of gastric carcinogenesis. Cancer Res. 1988 Jul 1;48(13):3554–3560. [PubMed] [Google Scholar]
  5. Crabtree J. E., Shallcross T. M., Heatley R. V., Wyatt J. I. Mucosal tumour necrosis factor alpha and interleukin-6 in patients with Helicobacter pylori associated gastritis. Gut. 1991 Dec;32(12):1473–1477. doi: 10.1136/gut.32.12.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cussac V., Ferrero R. L., Labigne A. Expression of Helicobacter pylori urease genes in Escherichia coli grown under nitrogen-limiting conditions. J Bacteriol. 1992 Apr;174(8):2466–2473. doi: 10.1128/jb.174.8.2466-2473.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Czinn S. J., Cai A., Nedrud J. G. Protection of germ-free mice from infection by Helicobacter felis after active oral or passive IgA immunization. Vaccine. 1993;11(6):637–642. doi: 10.1016/0264-410x(93)90309-l. [DOI] [PubMed] [Google Scholar]
  8. Eaton K. A., Krakowka S. Effect of gastric pH on urease-dependent colonization of gnotobiotic piglets by Helicobacter pylori. Infect Immun. 1994 Sep;62(9):3604–3607. doi: 10.1128/iai.62.9.3604-3607.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Engstrand L., Scheynius A., Påhlson C., Grimelius L., Schwan A., Gustavsson S. Association of Campylobacter pylori with induced expression of class II transplantation antigens on gastric epithelial cells. Infect Immun. 1989 Mar;57(3):827–832. doi: 10.1128/iai.57.3.827-832.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Ferrero R. L., Labigne A. Cloning, expression and sequencing of Helicobacter felis urease genes. Mol Microbiol. 1993 Jul;9(2):323–333. doi: 10.1111/j.1365-2958.1993.tb01693.x. [DOI] [PubMed] [Google Scholar]
  12. Fox J. G., Blanco M., Murphy J. C., Taylor N. S., Lee A., Kabok Z., Pappo J. Local and systemic immune responses in murine Helicobacter felis active chronic gastritis. Infect Immun. 1993 Jun;61(6):2309–2315. doi: 10.1128/iai.61.6.2309-2315.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fox J. G., Correa P., Taylor N. S., Lee A., Otto G., Murphy J. C., Rose R. Helicobacter mustelae-associated gastritis in ferrets. An animal model of Helicobacter pylori gastritis in humans. Gastroenterology. 1990 Aug;99(2):352–361. doi: 10.1016/0016-5085(90)91016-y. [DOI] [PubMed] [Google Scholar]
  14. Fox J. G., Lee A., Otto G., Taylor N. S., Murphy J. C. Helicobacter felis gastritis in gnotobiotic rats: an animal model of Helicobacter pylori gastritis. Infect Immun. 1991 Mar;59(3):785–791. doi: 10.1128/iai.59.3.785-791.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gootz T. D., Perez-Perez G. I., Clancy J., Martin B. A., Tait-Kamradt A., Blaser M. J. Immunological and molecular characterization of Helicobacter felis urease. Infect Immun. 1994 Mar;62(3):793–798. doi: 10.1128/iai.62.3.793-798.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Karttunen R. Blood lymphocyte proliferation, cytokine secretion and appearance of T cells with activation surface markers in cultures with Helicobacter pylori. Comparison of the responses of subjects with and without antibodies to H. pylori. Clin Exp Immunol. 1991 Mar;83(3):396–400. doi: 10.1111/j.1365-2249.1991.tb05650.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Labigne A., Cussac V., Courcoux P. Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity. J Bacteriol. 1991 Mar;173(6):1920–1931. doi: 10.1128/jb.173.6.1920-1931.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee A., Chen M., Coltro N., O'Rourke J., Hazell S., Hu P., Li Y. Long term infection of the gastric mucosa with Helicobacter species does induce atrophic gastritis in an animal model of Helicobacter pylori infection. Zentralbl Bakteriol. 1993 Sep;280(1-2):38–50. doi: 10.1016/s0934-8840(11)80939-4. [DOI] [PubMed] [Google Scholar]
  19. Lee A., Fox J. G., Otto G., Murphy J. A small animal model of human Helicobacter pylori active chronic gastritis. Gastroenterology. 1990 Nov;99(5):1315–1323. doi: 10.1016/0016-5085(90)91156-z. [DOI] [PubMed] [Google Scholar]
  20. Lee A., Hazell S. L., O'Rourke J., Kouprach S. Isolation of a spiral-shaped bacterium from the cat stomach. Infect Immun. 1988 Nov;56(11):2843–2850. doi: 10.1128/iai.56.11.2843-2850.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Leon-Barua R., Recavarren-Arce S., Gilman R. H., Berendson R. Can eradication of Helicobacter pylori prevent gastric cancer? Gastrointestinal Physiology Working Group of Cayetano Heredia. Drugs. 1993 Sep;46(3):341–346. doi: 10.2165/00003495-199346030-00001. [DOI] [PubMed] [Google Scholar]
  22. Marshall B. J., Warren J. R. Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet. 1984 Jun 16;1(8390):1311–1315. doi: 10.1016/s0140-6736(84)91816-6. [DOI] [PubMed] [Google Scholar]
  23. McGhee J. R., Kiyono H. New perspectives in vaccine development: mucosal immunity to infections. Infect Agents Dis. 1993 Apr;2(2):55–73. [PubMed] [Google Scholar]
  24. Nomura A., Stemmermann G. N., Chyou P. H., Kato I., Perez-Perez G. I., Blaser M. J. Helicobacter pylori infection and gastric carcinoma among Japanese Americans in Hawaii. N Engl J Med. 1991 Oct 17;325(16):1132–1136. doi: 10.1056/NEJM199110173251604. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Perez-Perez G. I., Dworkin B. M., Chodos J. E., Blaser M. J. Campylobacter pylori antibodies in humans. Ann Intern Med. 1988 Jul 1;109(1):11–17. doi: 10.7326/0003-4819-109-1-11. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Turbett G. R., Høj P. B., Horne R., Mee B. J. Purification and characterization of the urease enzymes of Helicobacter species from humans and animals. Infect Immun. 1992 Dec;60(12):5259–5266. doi: 10.1128/iai.60.12.5259-5266.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES