Immunoelectron microscopic localization of chlamydial lipopolysaccharide (LPS) in McCoy cells inoculated with Chlamydia trachomatis.
Interactions between Chlamydia trachomatis, host cells, and the immune system are believed to involve lipopolysaccharide (LPS). We used immunogold techniques to study the distribution of chlamydial LPS in cultured cells infected with C. trachomatis LGV-L1. McCoy cells inoculated with C. trachomatis were cultured and then fixed and embedded in situ with acrylic resins. Sections were immunolabeled with a protein A-gold method using antisera to the genus-specific, periodate-sensitive epitope on chlamydial LPS. Pre-embedding immunogold labeling on permeabilized cells was also done. By post-embedding methods, labeling for LPS was equally abundant over the outer membranes of elementary (EB) and reticulate bodies (RB). By post-embedding labeling, the sub-surface side of the EB outer membrane was more heavily labeled than the surface side. By pre-embedding labeling, LPS was found to be less abundant on the surface of EBs than RBs. Labeling for LPS was found over apparent lysosomes in McCoy cells and over electron-dense blebs on or near the surface of the plasma membranes of McCoy cells. These results indicate that the concentration of LPS in chlamydial membranes is constant during development but that with development its location changes from being mostly cell-surface to sub-surface. These results show that the post-embedding immunogold technique can be a useful approach for the cell culture-based study of chlamydial LPS.
Hearn
S A
SA
Department of Pathology, St Joseph's Health Center, London, Ontario, Canada.
McNabb
G L
GL
D000180:Acrylic Resins; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D002692:Chlamydia trachomatis / Q000254:growth & development / Q000276:immunology*; D007150:Immunohistochemistry; D002479:Inclusion Bodies / Q000276:immunology; D008070:Lipopolysaccharides / Q000032:analysis*; D008247:Lysosomes / Q000276:immunology; D051379:Mice; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D015388:Organelles / Q000276:immunology
D000180:Acrylic Resins; D008070:Lipopolysaccharides; C037883:Lowicryl 4KM resin; C048707:LR white