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. 1992 Oct;60(10):4433–4438. doi: 10.1128/iai.60.10.4433-4438.1992

Staining of surface antigens of Chlamydia trachomatis L2 in tissue culture.

M Baumann 1, L Brade 1, E Fasske 1, H Brade 1
PMCID: PMC257486  PMID: 1398957

Abstract

Surface labeling of chlamydial elementary and reticulate bodies in L929 cells infected with Chlamydia trachomatis serotype L2 was monitored by using monoclonal antibodies (MAb) against the major outer membrane protein and lipopolysaccharide (LPS). Different staining and fixation procedures were used to detect these surface antigens during the developmental cycle. Anti-major outer membrane protein MAb yielded a clear staining pattern of exclusively chlamydial inclusions independent of the fixation or staining technique used. Anti-LPS MAb gave a faint staining pattern of reticulate bodies when methanol fixation was used and showed that LPS was released from chlamydiae into the host cell cytoplasm and into the surroundings of the infected host cell. However, when paraformaldehyde-glutardialdehyde fixation was used, extracellular LPS staining was not observed. The data show that chlamydial LPS is loosely bound in the bacterial outer membrane but suggest that shedding of LPS is a fixation artifact.

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Selected References

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

  1. Baehr W., Zhang Y. X., Joseph T., Su H., Nano F. E., Everett K. D., Caldwell H. D. Mapping antigenic domains expressed by Chlamydia trachomatis major outer membrane protein genes. Proc Natl Acad Sci U S A. 1988 Jun;85(11):4000–4004. doi: 10.1073/pnas.85.11.4000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birkelund S., Lundemose A. G., Christiansen G. Chemical cross-linking of Chlamydia trachomatis. Infect Immun. 1988 Mar;56(3):654–659. doi: 10.1128/iai.56.3.654-659.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birkelund S., Lundemose A. G., Christiansen G. Immunoelectron microscopy of lipopolysaccharide in Chlamydia trachomatis. Infect Immun. 1989 Oct;57(10):3250–3253. doi: 10.1128/iai.57.10.3250-3253.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brade H., Brade L., Nano F. E. Chemical and serological investigations on the genus-specific lipopolysaccharide epitope of Chlamydia. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2508–2512. doi: 10.1073/pnas.84.8.2508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brade L., Holst O., Kosma P., Zhang Y. X., Paulsen H., Krausse R., Brade H. Characterization of murine monoclonal and murine, rabbit, and human polyclonal antibodies against chlamydial lipopolysaccharide. Infect Immun. 1990 Jan;58(1):205–213. doi: 10.1128/iai.58.1.205-213.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brade L., Schramek S., Schade U., Brade H. Chemical, biological, and immunochemical properties of the Chlamydia psittaci lipopolysaccharide. Infect Immun. 1986 Nov;54(2):568–574. doi: 10.1128/iai.54.2.568-574.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caldwell H. D., Hitchcock P. J. Monoclonal antibody against a genus-specific antigen of Chlamydia species: location of the epitope on chlamydial lipopolysaccharide. Infect Immun. 1984 May;44(2):306–314. doi: 10.1128/iai.44.2.306-314.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dutilh B., Bébéar C., Taylor-Robinson D., Grimont P. A. Detection of Chlamydia trachomatis by in situ hybridization with sulphonated total DNA. Ann Inst Pasteur Microbiol. 1988 Jan-Feb;139(1):115–127. doi: 10.1016/0769-2609(88)90099-3. [DOI] [PubMed] [Google Scholar]
  9. Fu Y., Baumann M., Kosma P., Brade L., Brade H. A synthetic glycoconjugate representing the genus-specific epitope of chlamydial lipopolysaccharide exhibits the same specificity as its natural counterpart. Infect Immun. 1992 Apr;60(4):1314–1321. doi: 10.1128/iai.60.4.1314-1321.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hearn S. A., McNabb G. L. Immunoelectron microscopic localization of chlamydial lipopolysaccharide (LPS) in McCoy cells inoculated with Chlamydia trachomatis. J Histochem Cytochem. 1991 Aug;39(8):1067–1075. doi: 10.1177/39.8.1649854. [DOI] [PubMed] [Google Scholar]
  11. Holst O., Brade L., Kosma P., Brade H. Structure, serological specificity, and synthesis of artificial glycoconjugates representing the genus-specific lipopolysaccharide epitope of Chlamydia spp. J Bacteriol. 1991 Mar;173(6):1862–1866. doi: 10.1128/jb.173.6.1862-1866.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Karimi S. T., Schloemer R. H., Wilde C. E., 3rd Accumulation of chlamydial lipopolysaccharide antigen in the plasma membranes of infected cells. Infect Immun. 1989 Jun;57(6):1780–1785. doi: 10.1128/iai.57.6.1780-1785.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kosma P., Bahnmüller R., Schulz G., Brade H. Synthesis of a tetrasaccharide of the genus-specific lipopolysaccharide epitope of Chlamydia. Carbohydr Res. 1990 Dec 15;208:37–50. doi: 10.1016/0008-6215(90)80083-f. [DOI] [PubMed] [Google Scholar]
  14. Kuo C. C., Chen H. H., Wang S. P., Grayston J. T. Identification of a new group of Chlamydia psittaci strains called TWAR. J Clin Microbiol. 1986 Dec;24(6):1034–1037. doi: 10.1128/jcm.24.6.1034-1037.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nano F. E., Caldwell H. D. Expression of the chlamydial genus-specific lipopolysaccharide epitope in Escherichia coli. Science. 1985 May 10;228(4700):742–744. doi: 10.1126/science.2581315. [DOI] [PubMed] [Google Scholar]
  16. Nurminen M., Leinonen M., Saikku P., Mäkelä P. H. The genus-specific antigen of Chlamydia: resemblance to the lipopolysaccharide of enteric bacteria. Science. 1983 Jun 17;220(4603):1279–1281. doi: 10.1126/science.6344216. [DOI] [PubMed] [Google Scholar]
  17. Nurminen M., Rietschel E. T., Brade H. Chemical characterization of Chlamydia trachomatis lipopolysaccharide. Infect Immun. 1985 May;48(2):573–575. doi: 10.1128/iai.48.2.573-575.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Richmond S. J., Stirling P. Localization of chlamydial group Antigen in McCoy cell monolayers infected with Chlamydia trachomatis or Chlamydia psittaci. Infect Immun. 1981 Nov;34(2):561–570. doi: 10.1128/iai.34.2.561-570.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schachter J. The intracellular life of Chlamydia. Curr Top Microbiol Immunol. 1988;138:109–139. [PubMed] [Google Scholar]
  20. Stephens R. S., Tam M. R., Kuo C. C., Nowinski R. C. Monoclonal antibodies to Chlamydia trachomatis: antibody specificities and antigen characterization. J Immunol. 1982 Mar;128(3):1083–1089. [PubMed] [Google Scholar]
  21. Stuart E. S., Tirrell S. M., MacDonald A. B. Characterization of an antigen secreted by Chlamydia-infected cell culture. Immunology. 1987 Aug;61(4):527–533. [PMC free article] [PubMed] [Google Scholar]
  22. Taylor H. R., Prendergast R. A. Attempted oral immunization with chlamydial lipopolysaccharide subunit vaccine. Invest Ophthalmol Vis Sci. 1987 Oct;28(10):1722–1726. [PubMed] [Google Scholar]
  23. Yong E. C., Chinn J. S., Caldwell H. D., Kuo C. C. Reticulate bodies as single antigen in Chlamydia trachomatis serology with microimmunofluorescence. J Clin Microbiol. 1979 Sep;10(3):351–356. doi: 10.1128/jcm.10.3.351-356.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Zhang Y. X., Stewart S., Joseph T., Taylor H. R., Caldwell H. D. Protective monoclonal antibodies recognize epitopes located on the major outer membrane protein of Chlamydia trachomatis. J Immunol. 1987 Jan 15;138(2):575–581. [PubMed] [Google Scholar]

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