title: Trafficking of Streptococcus uberis in bovine mammary epithelial cells.
abstract: Results from our laboratory showed that Streptococcus uberis internalized bovine mammary epithelial cells by exploiting host cell cytoskeleton and signal transduction mechanisms. It was also shown that S. uberis survived intracellularly for up to 120 h and capable of transcytose bovine mammary epithelial cells. To define mechanisms and strategies used by S. uberis to move through host cells and survive intracellularly, internalization studies using specific inhibitors, double immunofluorescence labeling and confocal laser microscopy were conducted. When bovine mammary epithelial cells were treated with inhibitors of endocytic vesicle acidification, the number of intracellular S. uberis was similar to untreated controls. When selective inhibitors of lipid rafts/caveolae or receptor-mediated endocytosis were used, a significantly lower number of intracellular S. uberis was detected compared with untreated controls. However, when the effect of inhibitors of receptor-mediated endocytosis and lipid rafts/caveolae were compared, the latter induced the lowest S. uberis internalization values suggesting a preferential exploitation of caveolae-mediated endocytosis. Since caveloae-dependent intracellular trafficking does not include intravesicular acidification or lysosome fusion; these results suggest that by exploiting preferential intracellular trafficking pathways in bovine mammary epithelial cells, S. uberis avoids intracellular bactericidal mechanisms. Such a strategy would allow S. uberis to persist intracellularly and may explain how persistent intramammary infections occur.
authors.affiliation: Department of Animal Science, The Food Safety Center of Excellence, Institute of Agriculture, The University of Tennessee, 60 MCCord Hall, Knoxville, TN 37996, USA. ralmeida@utk.edu
authors.forename: Raul A
authors.initials: RA
authors.lastname: Almeida
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Oliver
mesh_terms: D000818:Animals; D002417:Cattle; D021941:Caveolae / Q000187:drug effects / Q000382:microbiology / Q000502:physiology; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology; D011992:Endosomes / Q000378:metabolism / Q000382:microbiology; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology; D005260:Female; D005372:Filipin / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008321:Mammary Glands, Animal / Q000382:microbiology*; D043265:Phosphatidylinositol Diacylglycerol-Lyase / Q000494:pharmacology; D013290:Streptococcal Infections / Q000382:microbiology* / Q000662:veterinary*; D013291:Streptococcus / Q000378:metabolism / Q000502:physiology*; D047392:beta-Cyclodextrins / Q000494:pharmacology
chemical_list: D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D005372:Filipin; D043265:Phosphatidylinositol Diacylglycerol-Lyase