title: Association of caveolin with Chlamydia trachomatis inclusions at early and late stages of infection.
abstract: The mechanism by which the intracellular bacterial pathogen Chlamydia trachomatis enters eukaryotic cells is poorly understood. There are conflicting reports of entry occurring by clathrin-dependent and clathrin-independent processes. We report here that C. trachomatis serovar K enters HEp-2 and HeLa 229 epithelial cells and J-774A.1 mouse macrophage/monocyte cells via caveolin-containing sphingolipid and cholesterol-enriched raft microdomains in the host cell plasma membranes. First, filipin and nystatin, drugs that specifically disrupt raft function by cholesterol chelation, each impaired entry of C. trachomatis serovar K. In control experiments, filipin did not impair entry of the same organism by an antibody-mediated opsonic process, nor did it impair entry of BSA-coated microspheres. Second, the chlamydia-containing endocytic vesicles specifically reacted with antisera against the caveolae marker protein caveolin. These vesicles are known to become the inclusions in which parasite replication occurs. They avoid fusion with lysosomes and instead traffic to the Golgi region, where they intercept Golgi-derived vesicles that recycle sphingolipids and cholesterol to the plasma membrane. We also report that late-stage C. trachomatis inclusions continue to display high levels of caveolin, which they likely acquire from the exocytic Golgi vesicles. We suggest that the atypical raft-mediated entry process may have important consequences for the host-pathogen interaction well after entry has occurred. These consequences include enabling the chlamydial vesicle to avoid acidification and fusion with lysosomes, to traffic to the Golgi region, and to intercept sphingolipid-containing vesicles from the Golgi.
authors.affiliation: Department of Microbiology, University of Massachusetts, Amherst, Massachusetts 01003, USA. lnorkin@microbio.umass.edu
authors.forename: L C
authors.initials: LC
authors.lastname: Norkin
authors.forename: S A
authors.initials: SA
authors.lastname: Wolfrom
authors.forename: E S
authors.initials: ES
authors.lastname: Stuart
mesh_terms: D000818:Animals; D051242:Caveolin 1; D022461:Caveolins / Q000032:analysis*; D002460:Cell Line; D002692:Chlamydia trachomatis / Q000472:pathogenicity*; D004848:Epithelium / Q000382:microbiology; D005372:Filipin / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry / Q000382:microbiology*; D008264:Macrophages / Q000382:microbiology; D021962:Membrane Microdomains / Q000382:microbiology; D051379:Mice; D009761:Nystatin / Q000494:pharmacology; D010588:Phagosomes / Q000737:chemistry / Q000382:microbiology
chemical_list: C495540:CAV1 protein, human; C495541:Cav1 protein, mouse; D051242:Caveolin 1; D022461:Caveolins; D009761:Nystatin; D005372:Filipin