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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Jul;30(7):1743–1751. doi: 10.1128/jcm.30.7.1743-1751.1992

Immunoglobulin G antibodies to Helicobacter pylori in patients with dyspeptic symptoms investigated by the western immunoblot technique.

L P Andersen 1, F Espersen 1
PMCID: PMC265374  PMID: 1629330

Abstract

Helicobacter pylori is a gram-negative, curved, rod-shaped bacterium known to cause gastritis and to be an important factor in the pathogenesis of peptic ulcers. Serological testing has recently been proposed as an aid in diagnosis of H. pylori infections. In this study, we used the Western immunoblot technique to evaluate the possibility of using one or more of the antigens from H. pylori for this purpose. Thirteen major bands and about 30 minor bands could be identified by Western blotting when sera from 53 consecutive dyspeptic patients, 27 healthy children, and 25 blood donors were evaluated. Antibodies against most of the major bands were found significantly more frequently in patients with H. pylori infections than in patients without such infections. However, antibodies against a single polypeptide were not produced by all patients with H. pylori infection. Polypeptides with molecular masses of 120, 50, and between 19 and 36 kDa seem to be the most specific polypeptides for the diagnosis of H. pylori infections. This study showed only minor differences in antigenic composition between different clinical isolates of H. pylori, and serological cross-reactions with other bacteria were limited. Major serological cross-reactions were found only with Campylobacter jejuni and with bacterial lipopolysaccharide. However, one band at 60 kDa reacted with antiserum to the Legionella micdadei common antigen, which may indicate a cross-reaction with common antigen from several other bacteria. This study demonstrates that a number of bands may be useful as antigens in serological tests after isolation and purification.

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Selected References

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  1. Andersen L. P., Elsborg L., Justesen T. Campylobacter pylori in peptic ulcer disease. III. Symptoms and paraclinical and epidemiologic findings. Scand J Gastroenterol. 1988 Apr;23(3):347–350. doi: 10.3109/00365528809093877. [DOI] [PubMed] [Google Scholar]
  2. Andersen L. P., Holck S., Povlsen C. O. Campylobacter pylori detected by indirect immunohistochemical technique. APMIS. 1988 Jun;96(6):559–564. [PubMed] [Google Scholar]
  3. Andersen L. P., Holck S., Povlsen C. O., Elsborg L., Justesen T. Campylobacter pyloridis in peptic ulcer disease. I. Gastric and duodenal infection caused by C. pyloridis: histopathologic and microbiologic findings. Scand J Gastroenterol. 1987 Mar;22(2):219–224. doi: 10.3109/00365528708991883. [DOI] [PubMed] [Google Scholar]
  4. Bangsborg J. M., Collins M. T., Høiby N., Hindersson P. Cloning and expression of the Legionella micdadei "common antigen" in Escherichia coli. APMIS. 1989 Jan;97(1):14–22. [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Czinn S., Carr H., Sheffler L., Aronoff S. Serum IgG antibody to the outer membrane proteins of Campylobacter pylori in children with gastroduodenal disease. J Infect Dis. 1989 Mar;159(3):586–589. doi: 10.1093/infdis/159.3.586. [DOI] [PubMed] [Google Scholar]
  8. Espersen F., Clemmensen I., Barkholt V. Isolation of Staphylococcus aureus clumping factor. Infect Immun. 1985 Sep;49(3):700–708. doi: 10.1128/iai.49.3.700-708.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ferrero R. L., Hazell S. L., Lee A. The urease enzymes of Campylobacter pylori and a related bacterium. J Med Microbiol. 1988 Sep;27(1):33–40. doi: 10.1099/00222615-27-1-33. [DOI] [PubMed] [Google Scholar]
  10. Fomsgaard A., Conrad R. S., Galanos C., Shand G. H., Høiby N. Comparative immunochemistry of lipopolysaccharides from typable and polyagglutinable Pseudomonas aeruginosa strains isolated from patients with cystic fibrosis. J Clin Microbiol. 1988 May;26(5):821–826. doi: 10.1128/jcm.26.5.821-826.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Gnarpe H., Unge P., Blomqvist C., Mäkitalo S. Campylobacter pylori in Swedish patients referred for gastroscopy. APMIS. 1988 Feb;96(2):128–132. [PubMed] [Google Scholar]
  13. Guruge J. L., Schalén C., Nilsson I., Ljungh A., Tyszkiewicz T., Wikander M., Wadström T. Detection of antibodies to Helicobacter pylori cell surface antigens. Scand J Infect Dis. 1990;22(4):457–465. doi: 10.3109/00365549009027078. [DOI] [PubMed] [Google Scholar]
  14. Hirschl A. M., Pletschette M., Hirschl M. H., Berger J., Stanek G., Rotter M. L. Comparison of different antigen preparations in an evaluation of the immune response to Campylobacter pylori. Eur J Clin Microbiol Infect Dis. 1988 Aug;7(4):570–575. doi: 10.1007/BF01962618. [DOI] [PubMed] [Google Scholar]
  15. Kaldor J., Tee W., Nicolacopolous C., Demirtzoglou K., Noonan D., Dwyer B. Immunoblot confirmation of immune response to Campylobacter pyloridis in patients with duodenal ulcers. Med J Aust. 1986 Aug 4;145(3-4):133–135. doi: 10.5694/j.1326-5377.1986.tb113771.x. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Mills S. D., Kurjanczyk L. A., Penner J. L. Identification of an antigen common to different species of the genus Campylobacter. J Clin Microbiol. 1988 Jul;26(7):1411–1413. doi: 10.1128/jcm.26.7.1411-1413.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mitchell H. M., Lee A., Berkowicz J., Borody T. The use of serology to diagnose active Campylobacter pylori infection. Med J Aust. 1988 Dec 5;149(11-12):604–609. doi: 10.5694/j.1326-5377.1988.tb120800.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Morris A., Nicholson G., Lloyd G., Haines D., Rogers A., Taylor D. Seroepidemiology of Campylobacter pyloridis. N Z Med J. 1986 Sep 10;99(809):657–659. [PubMed] [Google Scholar]
  21. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  22. Nedenskov-Sørensen P., Bjørneklett A., Fausa O., Bukholm G., Aase S., Jantzen E. Campylobacter pylori infection and its relation to chronic gastritis. An endoscopic, bacteriologic, and histomorphologic study. Scand J Gastroenterol. 1988 Sep;23(7):867–874. doi: 10.3109/00365528809090775. [DOI] [PubMed] [Google Scholar]
  23. Newell D. G. Identification of the outer membrane proteins of Campylobacter pyloridis and antigenic cross-reactivity between C. pyloridis and C. jejuni. J Gen Microbiol. 1987 Jan;133(1):163–170. doi: 10.1099/00221287-133-1-163. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Rauws E. A., Langenberg W., Houthoff H. J., Zanen H. C., Tytgat G. N. Campylobacter pyloridis-associated chronic active antral gastritis. A prospective study of its prevalence and the effects of antibacterial and antiulcer treatment. Gastroenterology. 1988 Jan;94(1):33–40. [PubMed] [Google Scholar]
  27. Reiff A., Jacobs E., Kist M. Seroepidemiological study of the immune response to Campylobacter pylori in potential risk groups. Eur J Clin Microbiol Infect Dis. 1989 Jul;8(7):592–596. doi: 10.1007/BF01968135. [DOI] [PubMed] [Google Scholar]
  28. Schaber E., Umlauft F., Stöffler G., Aigner F., Paulweber B., Sandhofer F. Indirect immunofluorescence test and enzyme-linked immunosorbent assay for detection of Campylobacter pylori. J Clin Microbiol. 1989 Feb;27(2):327–330. doi: 10.1128/jcm.27.2.327-330.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Surveyor I., Goodwin C. S., Mullan B. P., Geelhoed E., Warren J. R., Murray R. N., Waters T. E., Sanderson C. R. The 14C-urea breath-test for the detection of gastric Campylobacter pylori infection. Med J Aust. 1989 Oct 16;151(8):435–439. [PubMed] [Google Scholar]
  30. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  32. von Wulffen H., Grote H. J. Enzyme-linked immunosorbent assay for detection of immunoglobulin A and G antibodies to Campylobacter pylori. Eur J Clin Microbiol Infect Dis. 1988 Aug;7(4):559–565. doi: 10.1007/BF01962616. [DOI] [PubMed] [Google Scholar]
  33. von Wulffen H., Heesemann J., Bützow G. H., Löning T., Laufs R. Detection of Campylobacter pyloridis in patients with antrum gastritis and peptic ulcers by culture, complement fixation test, and immunoblot. J Clin Microbiol. 1986 Nov;24(5):716–720. doi: 10.1128/jcm.24.5.716-720.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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