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Forward Genetic Approaches in Chlamydia trachomatis
Published on: October 23, 2013
Lipopolysaccharide in cells infected by Chlamydia trachomatis
S Campbell1, S J Richmond, P S Yates
1Department of Obstetrics and Gynaecology, University Hospital of South Manchester, West Didsbury, UK.
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 Chlamydia trachomatis 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 125I-labelled anti-LPS monoclonal antibodies to air-dried or methanol-fixed monolayers respectively, increased for up to 3 d then declined.(ABSTRACT TRUNCATED AT 250 WORDS)
Insights
Chlamydial lipopolysaccharide (LPS) is expressed on the host cell surface during infection, potentially spreading to neighboring cells. This study confirms LPS presence on host cells, not just within inclusions, using immunofluorescence and radioimmunoassays.
Area of Science:
- Microbiology
- Cell Biology
- Immunology
Background:
- The expression of chlamydial lipopolysaccharide (LPS) in eukaryotic host cells remains a subject of debate.
- Understanding LPS localization is crucial for elucidating Chlamydia pathogenesis.
Purpose of the Study:
- To investigate the presence and distribution of chlamydial LPS on the surface of infected host cells.
- To examine the dynamics of LPS expression during the chlamydial developmental cycle and its potential transmission.
Main Methods:
- Experimental infection of McCoy cell line, human endometrial, and amniotic epithelial cells with Chlamydia trachomatis serotype E.
- Immunofluorescence microscopy of air-dried host cells to visualize LPS distribution.
- Radioimmunoassays to quantify LPS levels in infected cell monolayers.
Main Results:
- Chlamydial LPS was detected on the surface of infected host cells, particularly near the chlamydial inclusion.
- LPS was observed at points of contact between infected and uninfected neighboring cells, forming immunofluorescent plaques.
- Cell-surface-associated and inclusion-associated LPS levels increased for up to 3 days post-infection before declining.
Conclusions:
- This study provides evidence for the expression of chlamydial LPS on the host cell surface.
- LPS may be transmitted to adjacent cells, contributing to the spread of infection.
- The findings challenge previous assumptions about LPS localization during Chlamydia infection.
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