title: Invasion of cultured human epithelial cells by Klebsiella pneumoniae isolated from the urinary tract.
abstract: The mechanisms which enable entry into cultured human epithelial cells by Klebsiella pneumoniae were compared with those of Salmonella typhi Ty2. K. pneumoniae 3091, isolated from a urine sample of a patient with a urinary tract infection, invaded human epithelial cells from the bladder and ileocecum and persisted for days in vitro. Electron microscopic studies demonstrated that K. pneumoniae was always contained in endosomes. The internalization mechanism(s) triggered by K. pneumoniae was studied by invasion assays conducted with different inhibitors that act on prokaryotic and eukaryotic cell structures and processes. Chloramphenicol inhibition of bacterial uptake revealed that bacterial de novo protein synthesis was essential for efficient invasion by K. pneumoniae and S. typhi. Interference with receptor-mediated endocytosis by g-strophanthin or monodansylcadaverine and inhibition of endosome acidification by monensin reduced the number of viable intracellular K. pneumoniae cells, but not S. typhi cells. The depolymerization of microfilaments by cytochalasin D inhibited the uptake of both bacteria. Microtubule depolymerization caused by colchicine, demecolcine, or nocodazole and the stabilization of microtubules with taxol reduced only the invasion ability of K. pneumoniae. S. typhi invasion was unaffected by microtubule depolymerization or stabilization. These data suggest that the internalization mechanism triggered by K. pneumoniae 3091 is strikingly different from the solely microfilament-dependent invasion mechanism exhibited by many of the well-studied enteric bacteria, such as enteroinvasive Escherichia coli, Salmonella, Shigella, and Yersinia strains.
authors.affiliation: Department of Bacterial Immunology, Walter Reed Army Institute of Research, Washington, D.C. 20307, USA.
authors.forename: T A
authors.initials: TA
authors.lastname: Oelschlaeger
authors.forename: B D
authors.initials: BD
authors.lastname: Tall
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis; D002103:Cadaverine / Q000031:analogs & derivatives / Q000494:pharmacology; D002478:Cells, Cultured; D002701:Chloramphenicol / Q000494:pharmacology; D003078:Colchicine / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D003703:Demecolcine / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects; D004847:Epithelial Cells; D004848:Epithelium / Q000382:microbiology; D006801:Humans; D007476:Ionophores / Q000494:pharmacology; D007711:Klebsiella pneumoniae / Q000472:pathogenicity / Q000502:physiology*; D008870:Microtubules / Q000187:drug effects; D008985:Monensin / Q000494:pharmacology; D015739:Nocodazole / Q000494:pharmacology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010042:Ouabain / Q000494:pharmacology; D017239:Paclitaxel / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012485:Salmonella typhi / Q000472:pathogenicity / Q000502:physiology; D014551:Urinary Tract / Q000382:microbiology*; D014552:Urinary Tract Infections / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D007476:Ionophores; D019384:Nucleic Acid Synthesis Inhibitors; D011500:Protein Synthesis Inhibitors; D015638:Cytochalasin D; D010042:Ouabain; D002701:Chloramphenicol; D008985:Monensin; C008542:monodansylcadaverine; D002103:Cadaverine; D017239:Paclitaxel; D015739:Nocodazole; D003078:Colchicine; D003703:Demecolcine