Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1985 Oct;50(1):284–291. doi: 10.1128/iai.50.1.284-291.1985

Basis for serological heterogeneity of thermostable antigens of Campylobacter jejuni.

S D Mills, W C Bradbury, J L Penner
PMCID: PMC262169  PMID: 2412962

Abstract

Lipopolysaccharides (LPS) were extracted from eight strains of Campylobacter jejuni and purified by enzyme treatment to remove traces of RNA, DNA, and protein. This material was used to sensitize sheep erythrocytes for the passive hemagglutination assay that is presently used to serotype C. jejuni. The results confirmed that the thermostable antigen typing scheme is based on LPS (O) antigens. The LPS after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining was found to consist of a series of slow migrating bands which could not be eliminated by treatment with NaOH, urea, or EDTA. However, the use of LPS double labeled with 14C and 32P yielded evidence that the bands of high molecular weight were indeed aggregations of low-molecular-weight LPS molecules.

Full text

PDF
291

Images in this article

287Image
on p.287
287Image
on p.287
288Image
on p.288
288Image
on p.288
289Image
on p.289

Selected References

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

  1. Abbott J. D., Dale B., Eldridge J., Jones D. M., Sutcliffe E. M. Serotyping of Campylobacter jejuni/coli. J Clin Pathol. 1980 Aug;33(8):762–766. doi: 10.1136/jcp.33.8.762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blaser M. J., Penner J. L., Wells J. G. Diversity of serotypes in outbreaks of enteritis due to Campylobacter jejuni. J Infect Dis. 1982 Dec;146(6):826–826. doi: 10.1093/infdis/146.6.826. [DOI] [PubMed] [Google Scholar]
  3. Bokkenheuser V. Vibrio fetus infection in man: a serological test. Infect Immun. 1972 Feb;5(2):222–226. doi: 10.1128/iai.5.2.222-226.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buck G. E., Smith J. S., Parshall K. A. Composition of the antigenic material removed from Campylobacter jejuni by heat. J Clin Microbiol. 1984 Dec;20(6):1094–1098. doi: 10.1128/jcm.20.6.1094-1098.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DAVIES D. A., CRUMPTON M. J., MACPHERSON I. A., HUTCHISON A. M. The adsorption of bacterial polysaccharides by erythrocytes. Immunology. 1958 Apr;1(2):157–171. [PMC free article] [PubMed] [Google Scholar]
  6. Darveau R. P., Hancock R. E. Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains. J Bacteriol. 1983 Aug;155(2):831–838. doi: 10.1128/jb.155.2.831-838.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eskenazy M., Cohen R. Salmonella O antigen-coated red blood cells as a stable reagent for passive hemagglutination tests. J Immunol Methods. 1974 Dec;6(1-2):129–131. doi: 10.1016/0022-1759(74)90098-2. [DOI] [PubMed] [Google Scholar]
  8. Goldman R. C., Leive L. Heterogeneity of antigenic-side-chain length in lipopolysaccharide from Escherichia coli 0111 and Salmonella typhimurium LT2. Eur J Biochem. 1980;107(1):145–153. doi: 10.1111/j.1432-1033.1980.tb04635.x. [DOI] [PubMed] [Google Scholar]
  9. Hatefi Y., Hanstein W. G. Solubilization of particulate proteins and nonelectrolytes by chaotropic agents. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1129–1136. doi: 10.1073/pnas.62.4.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hämmerling U., Westphal O. Synthesis and use of O-stearoyl polysaccharides in passive hemagglutination and hemolysis. Eur J Biochem. 1967 Mar;1(1):46–50. doi: 10.1007/978-3-662-25813-2_9. [DOI] [PubMed] [Google Scholar]
  11. Hébert G. A., Hollis D. G., Weaver R. E., Steigerwalt A. G., McKinney R. M., Brenner D. J. Serogroups of Campylobacter jejuni, Campylobacter coli, and Campylobacter fetus defined by direct immunofluorescence. J Clin Microbiol. 1983 Mar;17(3):529–538. doi: 10.1128/jcm.17.3.529-538.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Inzana T. J. Electrophoretic heterogeneity and interstrain variation of the lipopolysaccharide of Haemophilus influenzae. J Infect Dis. 1983 Sep;148(3):492–499. doi: 10.1093/infdis/148.3.492. [DOI] [PubMed] [Google Scholar]
  13. Karkhanis Y. D., Zeltner J. Y., Jackson J. J., Carlo D. J. A new and improved microassay to determine 2-keto-3-deoxyoctonate in lipopolysaccharide of Gram-negative bacteria. Anal Biochem. 1978 Apr;85(2):595–601. doi: 10.1016/0003-2697(78)90260-9. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. 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]
  16. Lauwers S., Vlaes L., Butzler J. P. Campylobacter serotyping a epidemiology. Lancet. 1981 Jan 17;1(8212):158–159. doi: 10.1016/s0140-6736(81)90748-0. [DOI] [PubMed] [Google Scholar]
  17. Leive L., Shovlin V. K., Mergenhagen S. E. Physical, chemical, and immunological properties of lipopolysaccharide released from Escherichia coli by ethylenediaminetetraacetate. J Biol Chem. 1968 Dec 25;243(24):6384–6391. [PubMed] [Google Scholar]
  18. 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]
  19. Logan S. M., Trust T. J. Structural and antigenic heterogeneity of lipopolysaccharides of Campylobacter jejuni and Campylobacter coli. Infect Immun. 1984 Jul;45(1):210–216. doi: 10.1128/iai.45.1.210-216.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McMyne P. M., Penner J. L., Mathias R. G., Black W. A., Hennessy J. N. Serotyping of Campylobacter jejuni isolated from sporadic cases and outbreaks in British Columbia. J Clin Microbiol. 1982 Aug;16(2):281–285. doi: 10.1128/jcm.16.2.281-285.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Mintz C. S., Apicella M. A., Morse S. A. Electrophoretic and serological characterization of the lipopolysaccharide produced by Neisseria gonorrhoeae. J Infect Dis. 1984 Apr;149(4):544–552. doi: 10.1093/infdis/149.4.544. [DOI] [PubMed] [Google Scholar]
  23. NETER E., WALKER J. Hemagglutination test for specific antibodies in dysentery caused by Shigella sonnei. Am J Clin Pathol. 1954 Dec;24(12):1424–1429. doi: 10.1093/ajcp/24.12_ts.1424. [DOI] [PubMed] [Google Scholar]
  24. NETER E., WESTPHAL O., LUDERITZ O., GORZYNSKI E. A., EICHENBERGER E. Studies of enterobacterial lipopolysaccharides; effects of heat and chemicals on erythrocyte-modifying, antigenic, toxic and pyrogenic properties. J Immunol. 1956 May;76(5):377–385. [PubMed] [Google Scholar]
  25. Naess V., Hofstad T. Chemical composition and biological activity of lipopolysaccharides prepared from type strains of Campylobacter jejuni and Campolybacter coli. Acta Pathol Microbiol Immunol Scand B. 1984 Aug;92(4):217–222. doi: 10.1111/j.1699-0463.1984.tb02824.x. [DOI] [PubMed] [Google Scholar]
  26. Naess V., Hofstad T. Chemical studies of partially hydrolysed lipopolysaccharides from four strains of Campylobacter jejuni and two strains of Campylobacter coli. J Gen Microbiol. 1984 Nov;130(11):2783–2789. doi: 10.1099/00221287-130-11-2783. [DOI] [PubMed] [Google Scholar]
  27. Naess V., Hofstad T. Isolation and chemical composition of lipopolysaccharide from Campylobacter jejuni. Acta Pathol Microbiol Immunol Scand B. 1982 Apr;90(2):135–139. doi: 10.1111/j.1699-0463.1982.tb00095.x. [DOI] [PubMed] [Google Scholar]
  28. Niwa M., Milner K. C., Ribi E., Rudbach J. A. Alteration of physical, chemical, and biological properties of endotoxin by treatment with mild alkali. J Bacteriol. 1969 Mar;97(3):1069–1077. doi: 10.1128/jb.97.3.1069-1077.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  30. Penner J. L., Hennessy J. N., Congi R. V. Serotyping of Campylobacter jejuni and Campylobacter coli on the basis of thermostable antigens. Eur J Clin Microbiol. 1983 Aug;2(4):378–383. doi: 10.1007/BF02019474. [DOI] [PubMed] [Google Scholar]
  31. Penner J. L., Hennessy J. N., Mills S. D., Bradbury W. C. Application of serotyping and chromosomal restriction endonuclease digest analysis in investigating a laboratory-acquired case of Campylobacter jejuni enteritis. J Clin Microbiol. 1983 Dec;18(6):1427–1428. doi: 10.1128/jcm.18.6.1427-1428.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Penner J. L., Pearson A. D., Hennessy J. N. Investigation of a waterborne outbreak of Campylobacter jejuni enteritis with a serotyping scheme based on thermostable antigens. J Clin Microbiol. 1983 Dec;18(6):1362–1365. doi: 10.1128/jcm.18.6.1362-1365.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Perez Perez G. I., Blaser M. J. Lipopolysaccharide characteristics of pathogenic campylobacters. Infect Immun. 1985 Feb;47(2):353–359. doi: 10.1128/iai.47.2.353-359.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schneider H., Hale T. L., Zollinger W. D., Seid R. C., Jr, Hammack C. A., Griffiss J. M. Heterogeneity of molecular size and antigenic expression within lipooligosaccharides of individual strains of Neisseria gonorrhoeae and Neisseria meningitidis. Infect Immun. 1984 Sep;45(3):544–549. doi: 10.1128/iai.45.3.544-549.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Skirrow M. B. Campylobacter enteritis - the first five years. J Hyg (Lond) 1982 Oct;89(2):175–184. doi: 10.1017/s0022172400070704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Smibert R. M. The genus Campylobacter. Annu Rev Microbiol. 1978;32:673–709. doi: 10.1146/annurev.mi.32.100178.003325. [DOI] [PubMed] [Google Scholar]
  38. Tsai C. M., Boykins R., Frasch C. E. Heterogeneity and variation among Neisseria meningitidis lipopolysaccharides. J Bacteriol. 1983 Aug;155(2):498–504. doi: 10.1128/jb.155.2.498-504.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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 Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES