1887

Abstract

In view of the controversy concerning the expression of chlamydial lipopolysaccharide (LPS) in the eukaryotic host cell, the presence of this molecule was examined in three cell types which were experimentally infected with serotype E. LPS was detected in the McCoy cell line, human endometrial epithelium and human amniotic epithelium with two monoclonal antibodies. The appearance and distribution of LPS at the host cell surface during the chlamydial developmental cycle and its transmission to neighbouring cells were examined by immunofluorescence microscopy after air drying of the host cells. LPS distribution was not uniform; it was first observed on regions of the cell surface in close proximity to the chlamydial endosome (inclusion). Soon after, the antigen was also detected at points of contact with neighbouring uninfected cells. Immunofluorescent plaques of host cells contaminated with LPS were thus formed in the vicinity of infected cells. These plaques increased in size over 2 d before becoming smaller as host cell lysis occurred. The major outer-membrane protein (MOMP) was not visualized on the host cell surface after air drying. No cell-surface LPS antigen was observed in live cells or those fixed in formaldehyde without air drying. Conventional methanol fixation and immunolocalization of LPS and MOMP in parallel infected cultures stained these antigens within inclusions in the expected fashion. Radio-immunoassays were used to quantify LPS in confluent McCoy cell monolayers during the chlamydial developmental cycle. Cell-surface-associated and inclusion-associated LPS, measured by direct binding of I-labelled anti-LPS monoclonal antibodies to air-dried or methanol-fixed monolayers respectively, increased for up to 3 d then declined. Cell-surface-associated LPS is not directly accessible to antibodies in the hydrated cell. This apparent masking of the antigen may have a significant advantage for persistence of the parasite , since such host-cell-associated antigen is unlikely to be a target for immune attack.

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1994-08-01
2021-07-30
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References

  1. Aplin J.D., Campbell S., Allen T.D. Human amnion extracellular matrix ultrastructure composition and deposition. J Cell Sci 1985; 79:119–136
    [Google Scholar]
  2. Bacon E.J., Richmond S.J., Wood D.J., Stirling P., Bevan B.J., Chalmers W.S.K. Serological detection of phage infection in Chlamydia psittaci recovered from ducks. I 'et Rec 1986; 119:618–620
    [Google Scholar]
  3. Baumann M., Brade L., Fasske E., Brade H. Staining of surface antigen of Chlamydia trachomatis L2 in tissue culture. Infect Immun 1992; 60:4433–4438
    [Google Scholar]
  4. Campbell S., Richmond S.J., Haynes P., Gump D.W., Yates P., Allen T.D. An in vitro model of Chlamydia trachomatis infection in the regenerative phase of the human endometrial cycle. J Gen Microbiol 1988; 134:2077–2088
    [Google Scholar]
  5. Campbell S., Richmond S.J., Yates P. The development of Chlamydia trachomatis inclusions within the host eukaryotic cell during interphase and mitosis. J Gen Microbiol 1989; 135:1153–1165
    [Google Scholar]
  6. Campbell S., Yates P.S., Waters F., Richmond S.J. Purification of Chlamydia trachomatis by a simple and rapid filtration method. J Gen Microbiol 1991; 137:1565–1569
    [Google Scholar]
  7. Dhir S.P., Kenney G.E., Grayson J.T. Characterisation of the group antigen of Chlamydia trachomatis. Infect Immun 1971; 4:725–730
    [Google Scholar]
  8. Dwyer R.C., Treharne J.D., Jones B.R., Herring J. Chlamydial infection. Results of microimmunofluorescence tests for the detection of type specific antibody in certain chlamydial infections. Br J Infect Dis 1972; 48:452–458
    [Google Scholar]
  9. Francis T., Magill T.P. An unidentified virus producing acute meningitis and pneumonitis in experimental animals. J Exp Med 1938; 68:147–160
    [Google Scholar]
  10. Galanos C., Luderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem 1969; 9:245–249
    [Google Scholar]
  11. Hearn S.A., McNabb G.L. Immunoelectron microscope localisation of chlamydial lipopolysaccharide (LPS) in McCoy cells inoculated with Chlamydia trachomatis. J Histochem Cytochem 1991; 39:1067–1075
    [Google Scholar]
  12. Howard L., Orenstein N.S., King N.W. Purification on renografin density gradients of Chlamydia trachomatis grown in the yolk sac of eggs. Appl Microbiol 1974; 27:102–106
    [Google Scholar]
  13. Igietseme J.U., Rank R.G. Susceptibility to reinfection after a primary chlamydial genital infection associated with a decrease of antigen-specific T cells in the genital tract. Infect Immun 1991; 59:1346–1351
    [Google Scholar]
  14. Karimi S.T., Schloemer R.H., Wilde C.E. Accumulation of chlamydial lipopolysaccharide antigen in the plasma membranes of infected cells. Infect Immun 1989; 57:1780–1785
    [Google Scholar]
  15. Mearns G., Richmond S.J., Storey C.C. Sensitive immune dot blot test for diagnosis of Chlamydia trachomatis. J Clin Microbiol 1988; 156:1810–1813
    [Google Scholar]
  16. Morgan H.R., Wiseman R.W. Growth of psittacosis virus in roller tube tissue culture; use in a vaccine. Infect Dis 1946; 79:131–133
    [Google Scholar]
  17. Morrison R.P. Immune responses to chlamydia are protective and pathogenic. In Proceedings of the 7th International Symposium on Human Chlamydial Infections 1990 Edited by Bowie W.R., Caldwell H.D., Jones R.P., Mardh P.A., Ridgway G.L., Schachter J., Stamm W.E., Ward M.E. Cambridge: Cambridge University Press; pp 163–172
    [Google Scholar]
  18. Murray E.S. Guinea pig inclusion conjunctivitis virus. 1. Isolation and identification as a member of the psittacosis-lymphogranuloma-trachoma group. J Infect Dis 1964; 114:1–12
    [Google Scholar]
  19. Pavia C.S., Schachter J. Failure to detect cell-mediated cytotoxicity against Chlamydia trachomatis-mlcctcd cells. Infect Immun 1983; 39:1271–1274
    [Google Scholar]
  20. Qvigstad E., Hirschberg H. Lack of cell-mediated cytotoxicity towards Chlamydia trachomatis infected target cells in humans. Acta Pathol Microbiol Immunol Scand Sect C 1984; 92:153–159
    [Google Scholar]
  21. Richmond S.J. Chlamydial group antigen in McCoy cells infected with Chlamydia trachomatis and Chlamydia psittaci. FEMS Microbiol Eett 1980; 8:47–50
    [Google Scholar]
  22. Richmond S.J., Stirling P. Localisation of chlamydial group antigen in McCoy cell monolayers infected with Chlamydia trachomatis or Chlamydia psittaci. Infect Immun 1981; 34:561–570
    [Google Scholar]
  23. Richmond S.J., Stirling P., Ashley C.R. Virus infecting the reticulate bodies of an avian strain of Chlamydia trachomatis. FEMS Microbiol Eett 1982; 14:31–36
    [Google Scholar]
  24. Richmond S.J., Storey C.C., Lusher M. Transmission of phage Chpl within Chlamydia. In Proceedings of the 7th International Symposium on Human Chlamydial Infections 1990 Edited by Bowie W.R., Caldwell H.D., Jones R.P., Mardh P.A., Ridgway G.L., Schachter J., Stamm W.E., Ward M.E. Cambridge: Cambridge University Press; pp 153–165
    [Google Scholar]
  25. Sompolinsky D., Richmond S.J. Growth of Chlamydia trachomatis in McCoy cells treated with cytochalasin B. Appl Microbiol 1974; 28:912–914
    [Google Scholar]
  26. Stagg A.J., Elsley W.A.J., Holland M., Ward M.E., Knight S.C. Dendritic cells (DC) in the initiation of immune responses to chlamydia. In Proceedings of the European Society for Chlamydial Research 1992 Edited by Mardh P.-A., La Placa M., Ward M. Uppsala: European Society fo; 2 pp 77–80
    [Google Scholar]
  27. Stirling P., Richmond S.J. The developmental cycle of Chlamydia trachomatis in McCoy cells treated with cytochalasin B. J Gen Microbiol 1977; 100:31–42
    [Google Scholar]
  28. Stirling P., Richmond S.J. Production of outer membrane blebs during chlamydial replication. FEMS Microbiol Lett 1980; 9:103–105
    [Google Scholar]
  29. Storey C.C., Mearns G., Richmond S.J. Immune dot blot technique for diagnosing infection with Chlamydia trachomatis. Genitourin Med 1987; 63:357–379
    [Google Scholar]
  30. Storey C., Lusher M., Yates P., Richmond S. Evidence for Chlamydiae pneumoniae of non-human origin. J Gen Microbiol 1993; 139:2621–2626
    [Google Scholar]
  31. Strauss J., Bednar B., Sery V. The incidence of ornithosis and salmonellosis in the black-headed gull. II. Isolation and identification of ornithosis virus in the gull and simultaneous detection of Salmonella typhimurium. Cesk Epidemiol 1957; 6:231–239
    [Google Scholar]
  32. Stuart E.S., Wyrick P.B., Choong J., Stoler S.B., Macdonald A.B. Examination of chlamydial glycolipid with monoclonal antibodies: cellular distribution and epitope binding. Immunology 1991; 74:740–747
    [Google Scholar]
  33. Thornley M.J., Zamze S.E., Byrne M.D., Lusher M., Evans R.T. Properties of monoclonal antibodies to the genus specific antigen of Chlamydia and their use for antigen detection by reverse passive haemagglutination. J Gen Microbiol 1985; 131:7–15
    [Google Scholar]
  34. Wang S.P., Grayston J.T. Classification of trachoma and related strains by microimmunofluorescence. In Trachoma and Related Disorders 1971 Edited by Nichols R.L. Amsterdam: Exerpta Medica; pp 305–321
    [Google Scholar]
  35. Wilde C.E., Karimi S.T., Haak R.A. Cell surface alterations during chlamydial infections. In Microbiology-1986 1986 Edited by Leive P.F., Boventre J.A., Morello J.A., Silver S.D., Wu H.C. Washington, DC: American Society for Microbiology; pp 96–98
    [Google Scholar]
  36. Wills J., Gruffydd-Jones T.J., Richmond S., Paul I.D. Isolation of Chlamydia psittaci from cases of conjunctivitis in a colony of cats. Vet Rec 1984; 114:344–346
    [Google Scholar]
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