title: The chlamydial inclusion preferentially intercepts basolaterally directed sphingomyelin-containing exocytic vacuoles.
abstract: Chlamydiae replicate intracellularly within a unique vacuole termed the inclusion. The inclusion circumvents classical endosomal/lysosomal pathways but actively intercepts a subset of Golgi-derived exocytic vesicles containing sphingomyelin (SM) and cholesterol. To further examine this interaction, we developed a polarized epithelial cell model to study vectoral trafficking of lipids and proteins to the inclusion. We examined seven epithelial cell lines for their ability to form single monolayers of polarized cells and support chlamydial development. Of these cell lines, polarized colonic mucosal C2BBe1 cells were readily infected with Chlamydia trachomatis and remained polarized throughout infection. Trafficking of (6-((N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl) amino)hexanoyl)sphingosine) (NBD-C(6)-ceramide) and its metabolic derivatives, NBD-glucosylceramide (GlcCer) and NBD-SM, was analyzed. SM was retained within L2-infected cells relative to mock-infected cells, correlating with a disruption of basolateral SM trafficking. There was no net retention of GlcCer within L2-infected cells and purification of C. trachomatis elementary bodies from polarized C2BBe1 cells confirmed that bacteria retained only SM. The chlamydial inclusion thus appears to preferentially intercept basolaterally-directed SM-containing exocytic vesicles, suggesting a divergence in SM and GlcCer trafficking. The observed changes in lipid trafficking were a chlamydia-specific effect because Coxiella burnetii-infected cells revealed no changes in GlcCer or SM polarized trafficking.
authors.affiliation: Host-Parasite Interactions Section, Laboratory of Intracellular Parasites, National Institute of Allergy and Infectious Diseases, Rocky Mountain Laboratories, 903 South 4th Street, Hamilton, Montana 59840, USA.
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Moore
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Fischer
authors.forename: David J
authors.initials: DJ
authors.lastname: Mead
authors.forename: Ted
authors.initials: T
authors.lastname: Hackstadt
mesh_terms: D001692:Biological Transport; D002460:Cell Line; D002692:Chlamydia trachomatis / Q000502:physiology*; D004847:Epithelial Cells / Q000648:ultrastructure; D005089:Exocytosis; D006801:Humans; D050356:Lipid Metabolism; D046529:Microscopy, Electron, Transmission; D014617:Vacuoles / Q000378:metabolism*
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