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. 1992 Nov;60(11):4826–4831. doi: 10.1128/iai.60.11.4826-4831.1992

Monoclonal antibodies against the native urease of Helicobacter pylori: synergistic inhibition of urease activity by monoclonal antibody combinations.

K Nagata 1, T Mizuta 1, Y Tonokatu 1, Y Fukuda 1, H Okamura 1, T Hayashi 1, T Shimoyama 1, T Tamura 1
PMCID: PMC258237  PMID: 1383158

Abstract

Monoclonal antibodies (MAbs) against the native urease of Helicobacter pylori NCTC 11637 were found to clearly inhibit the urease activity. Interestingly, synergistic inhibition by two MAbs recognizing different subunits was also observed. Ten MAbs were produced and classified as two isotypes of the immunoglobulin G (IgG) subclass, IgG1, and IgG2a. Western blot (immunoblot) analysis using sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that five MAbs recognized the large subunit and the other five recognized the small subunit of the urease. Among the MAbs, L2 and S2, which recognized the large and the small subunits, respectively, were also able to inhibit the urease activity of clinical isolates from H. pylori-infected patients. The combination of L2 and S2 led to augmented synergistic inhibition. L2, but not S2, could also inhibit the urease activity from Helicobacter mustelae; enzyme-linked immunosorbent assay and Western blot analysis showed that L2 cross-reacted with this urease. These results suggested that the epitope recognized by L2 had a structure common to both Helicobacter species and may be involved in the active site of the urease. In contrast to the MAbs, a polyclonal antibody in sera from mice immunized with H. pylori urease did not have the ability to inhibit H. pylori urease activity. However, the polyclonal antibody retained the ability to abolish the inhibitory action of these MAbs. Moreover, other MAbs which could not inhibit H. pylori urease activity also abolished the inhibitory action.

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Selected References

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  1. Barer M. R., Elliott T. S., Berkeley D., Thomas J. E., Eastham E. J. Cytopathic effects of Campylobacter pylori urease. J Clin Pathol. 1988 May;41(5):597–597. doi: 10.1136/jcp.41.5.597-a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bayerdörffer E., Oertel H., Lehn N., Kasper G., Mannes G. A., Sauerbruch T., Stolte M. Topographic association between active gastritis and Campylobacter pylori colonisation. J Clin Pathol. 1989 Aug;42(8):834–839. doi: 10.1136/jcp.42.8.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bode G., Malfertheiner P., Nilius M., Lehnhardt G., Ditschuneit H. Ultrastructural localisation of urease in outer membrane and periplasm of Campylobacter pylori. J Clin Pathol. 1989 Jul;42(7):778–779. doi: 10.1136/jcp.42.7.778-b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bolton F. J., Hutchinson D. N. Evaluation of three Campylobacter pylori antigen preparations for screening sera from patients undergoing endoscopy. J Clin Pathol. 1989 Jul;42(7):723–726. doi: 10.1136/jcp.42.7.723. [DOI] [PMC free article] [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. Coghlan J. G., Gilligan D., Humphries H., McKenna D., Dooley C., Sweeney E., Keane C., O'Morain C. Campylobacter pylori and recurrence of duodenal ulcers--a 12-month follow-up study. Lancet. 1987 Nov 14;2(8568):1109–1111. doi: 10.1016/s0140-6736(87)91545-5. [DOI] [PubMed] [Google Scholar]
  7. Dent J. C., McNulty C. A., Uff J. S., Gear M. W., Wilkinson S. P. Campylobacter pylori urease: a new serological test. Lancet. 1988 Apr 30;1(8592):1002–1002. doi: 10.1016/s0140-6736(88)91827-2. [DOI] [PubMed] [Google Scholar]
  8. Dunn B. E., Campbell G. P., Perez-Perez G. I., Blaser M. J. Purification and characterization of urease from Helicobacter pylori. J Biol Chem. 1990 Jun 5;265(16):9464–9469. [PubMed] [Google Scholar]
  9. Dunn B. E., Perez-Perez G. I., Blaser M. J. Two-dimensional gel electrophoresis and immunoblotting of Campylobacter pylori proteins. Infect Immun. 1989 Jun;57(6):1825–1833. doi: 10.1128/iai.57.6.1825-1833.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dunn B. E., Sung C. C., Taylor N. S., Fox J. G. Purification and characterization of Helicobacter mustelae urease. Infect Immun. 1991 Sep;59(9):3343–3345. doi: 10.1128/iai.59.9.3343-3345.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Graham D. Y. Campylobacter pylori and peptic ulcer disease. Gastroenterology. 1989 Feb;96(2 Pt 2 Suppl):615–625. doi: 10.1016/s0016-5085(89)80057-5. [DOI] [PubMed] [Google Scholar]
  12. Hawtin P. R., Stacey A. R., Newell D. G. Investigation of the structure and localization of the urease of Helicobacter pylori using monoclonal antibodies. J Gen Microbiol. 1990 Oct;136(10):1995–2000. doi: 10.1099/00221287-136-10-1995. [DOI] [PubMed] [Google Scholar]
  13. Hazell S. L., Lee A. Campylobacter pyloridis, urease, hydrogen ion back diffusion, and gastric ulcers. Lancet. 1986 Jul 5;2(8497):15–17. doi: 10.1016/s0140-6736(86)92561-4. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Inouye H., Mizuta T., Shimoyama T., Nagata K., Tanaka T., Tamura T. Acute lethality of Helicobacter pylori in mice. A novel heat-labile toxin. Gastroenterol Jpn. 1991 Apr;26(2):230–230. doi: 10.1007/BF02811086. [DOI] [PubMed] [Google Scholar]
  16. Inouye H., Yamamoto I., Tanida N., Mikami J., Tamura K., Ohno T., Kano M., Shimoyama T. Campylobacter pylori in Japan: bacteriological feature and prevalence in healthy subjects and patients with gastroduodenal disorders. Gastroenterol Jpn. 1989 Oct;24(5):494–504. doi: 10.1007/BF02773875. [DOI] [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. Markwell M. A., Haas S. M., Tolbert N. E., Bieber L. L. Protein determination in membrane and lipoprotein samples: manual and automated procedures. Methods Enzymol. 1981;72:296–303. doi: 10.1016/s0076-6879(81)72018-4. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Musgrove C., Bolton F. J., Krypczyk A. M., Temperley J. M., Cairns S. A., Owen W. G., Hutchinson D. N. Campylobacter pylori: clinical, histological, and serological studies. J Clin Pathol. 1988 Dec;41(12):1316–1321. doi: 10.1136/jcp.41.12.1316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Newell D. G., Stacey A. Antigens for the serodiagnosis of Campylobacter pylori infections. Gastroenterol Clin Biol. 1989;13(1 Pt 1):37B–41B. [PubMed] [Google Scholar]
  23. Pallen M. J., Clayton C. L. Vaccination against Helicobacter pylori urease. Lancet. 1990 Jul 21;336(8708):186–187. doi: 10.1016/0140-6736(90)91716-n. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Xu J. K., Goodwin C. S., Cooper M., Robinson J. Intracellular vacuolization caused by the urease of Helicobacter pylori. J Infect Dis. 1990 Jun;161(6):1302–1304. doi: 10.1093/infdis/161.6.1302. [DOI] [PubMed] [Google Scholar]

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