Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1985 Jan;21(1):33–38. doi: 10.1128/jcm.21.1.33-38.1985

Western blot analysis of the human antibody response to Campylobacter jejuni cellular antigens during gastrointestinal infection.

I Nachamkin, A M Hart
PMCID: PMC271575  PMID: 2578479

Abstract

Western blot analysis was used to identify antigenic components of Campylobacter jejuni whole cells and outer membranes that elicit antibody responses in patients with campylobacter enteritis. Acute- and convalescent-phase sera from eight patients were analyzed for antibody activity against their homologous infecting strains and heterologous clinical isolates. Whole-cell and Sarkosyl-insoluble membrane components were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred electrophoretically to nitrocellulose paper for immunoblotting experiments. After the separated components were probed with patient sera, antibody binding was detected by autoradiography with 125I-protein A. Using this method, we detected several immunogenic components in whole cells and outer membranes. In the acute-phase response of some patients to infection, two to three components with approximate molecular weights of 66,000 (p66), 43,000 to 46,000 (major outer membrane protein), and 12,000 (p12) were detected in immunoblots. Convalescent-phase sera showed a more broad array of antibody binding to cell components. p66, shown to be campylobacter flagellin, was the major immunodominant component in almost all sera tested, however, p66 was not a major protein in Coomassie blue-stained gels. The major outer-membrane protein also bound to antibody, but with less intensity than p66. In general, the antibody specificity of patient sera was not limited to the homologous infecting strain, and antibodies cross-reacted with most components in heterologous strains. A low-molecular-weight component, identified as lipopolysaccharide with a modified silver stain, showed serological specificity for some patient sera. The results of this study showed that the antibody response of patients with campylobacter enteritis to C. jejuni antigens is variable. Flagellin appeared to be the major immunodominant component during infection.

Full text

PDF
33

Images in this article

34Image
on p.34
35Image
on p.35
35Image
on p.35
36Image
on p.36
37Image
on p.37
37Image
on p.37

Selected References

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

  1. Blaser M. J., Hopkins J. A., Berka R. M., Vasil M. L., Wang W. L. Identification and characterization of Campylobacter jejuni outer membrane proteins. Infect Immun. 1983 Oct;42(1):276–284. doi: 10.1128/iai.42.1.276-284.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blaser M. J., Hopkins J. A., Vasil M. L. Campylobacter jejuni outer membrane proteins are antigenic for humans. Infect Immun. 1984 Mar;43(3):986–993. doi: 10.1128/iai.43.3.986-993.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blaser M. J., Wells J. G., Feldman R. A., Pollard R. A., Allen J. R. Campylobacter enteritis in the United States. A multicenter study. Ann Intern Med. 1983 Mar;98(3):360–365. doi: 10.7326/0003-4819-98-3-360. [DOI] [PubMed] [Google Scholar]
  4. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  5. Guerrant R. L., Lahita R. G., Winn W. C., Jr, Roberts R. B. Campylobacteriosis in man: pathogenic mechanisms and review of 91 bloodstream infections. Am J Med. 1978 Oct;65(4):584–592. doi: 10.1016/0002-9343(78)90845-8. [DOI] [PubMed] [Google Scholar]
  6. Kaldor J., Pritchard H., Serpell A., Metcalf W. Serum antibodies in Campylobacter enteritis. J Clin Microbiol. 1983 Jul;18(1):1–4. doi: 10.1128/jcm.18.1.1-4.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kosunen T. U., Rautelin H., Pitkänen T., Pönkä A., Pettersson T. Antibodies against an acid extract from a single campylobacter strain in hospitalized campylobacter patients. Infection. 1983 Jul-Aug;11(4):189–191. doi: 10.1007/BF01641193. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Lambert M. E., Schofield P. F., Ironside A. G., Mandal B. K. Campylobacter colitis. Br Med J. 1979 Mar 31;1(6167):857–859. doi: 10.1136/bmj.1.6167.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lior H., Woodward D. L., Edgar J. A., Laroche L. J., Gill P. Serotyping of Campylobacter jejuni by slide agglutination based on heat-labile antigenic factors. J Clin Microbiol. 1982 May;15(5):761–768. doi: 10.1128/jcm.15.5.761-768.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Logan S. M., Trust T. J. Molecular identification of surface protein antigens of Campylobacter jejuni. Infect Immun. 1983 Nov;42(2):675–682. doi: 10.1128/iai.42.2.675-682.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Logan S. M., Trust T. J. Outer membrane characteristics of Campylobacter jejuni. Infect Immun. 1982 Dec;38(3):898–906. doi: 10.1128/iai.38.3.898-906.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mills S. D., Bradbury W. C. Human antibody response to outer membrane proteins of Campylobacter jejuni during infection. Infect Immun. 1984 Feb;43(2):739–743. doi: 10.1128/iai.43.2.739-743.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Penner J. L., Hennessy J. N. Passive hemagglutination technique for serotyping Campylobacter fetus subsp. jejuni on the basis of soluble heat-stable antigens. J Clin Microbiol. 1980 Dec;12(6):732–737. doi: 10.1128/jcm.12.6.732-737.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Price A. B., Jewkes J., Sanderson P. J. Acute diarrhoea: Campylobacter colitis and the role of rectal biopsy. J Clin Pathol. 1979 Oct;32(10):990–997. doi: 10.1136/jcp.32.10.990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ruiz-Palacios G. M., Torres J., Torres N. I., Escamilla E., Ruiz-Palacios B. R., Tamayo J. Cholera-like enterotoxin produced by Campylobacter jejuni. Characterisation and clinical significance. Lancet. 1983 Jul 30;2(8344):250–253. doi: 10.1016/s0140-6736(83)90234-9. [DOI] [PubMed] [Google Scholar]
  17. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES