title: Cell invasion and survival of Shiga toxin-producing Escherichia coli within cultured human intestinal epithelial cells.
abstract: Shiga toxin-producing Escherichia coli (STEC) cause severe human infections and their virulence abilities are not fully understood. Cattle are a key reservoir, and the terminal rectum is the principal site of bacterial carriage. Most STEC possess a pathogenicity island termed the locus of enterocyte effacement (LEE). Nonetheless, LEE-negative STEC have been associated with disease. We found that invasion of LEE-positive and LEE-negative strains was higher for human enterocytic cell lines and for undifferentiated Caco-2 cells. Intracellular bacteria could be detected as early as 5 min after infection and transmission electron microscopy showed bacteria within membrane-bound vacuoles. STEC invasion depended on actin microfilaments and protein kinases. Scanning electron microscopy revealed that bacterial entry was not associated with membrane ruffling. Absence of macropinocytosis or actin rearrangement at the entry points suggests a zipper-like entry mechanism. Disruption of the tight junction by EGTA enhanced invasion of Caco-2 monolayers, and bacterial invasion mostly proceeded through the basolateral pole of enterocytes. STEC persisted within Caco-2 cells for up to 96 h without cell death and bacterial viability increased after 48 h, suggesting intracellular multiplication. The relatively harmless intracellular localization of STEC can be an efficient strategy to prevent its elimination from the bovine intestinal tract.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro 20551-030, Brazil.
authors.forename: Fabiana
authors.initials: F
authors.lastname: Cordeiro
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro 20551-030, Brazil.
authors.forename: Rita Ifuoe K
authors.initials: RIK
authors.lastname: da Silva
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro 20551-030, Brazil.
authors.forename: Thaís L Z
authors.initials: TLZ
authors.lastname: Vargas-Stampe
authors.affiliation: Departamento de Microbiologia e Parasitologia, Universidade Federal Fluminense, Niterói 24210-130, Brazil.
authors.forename: Aloysio M F
authors.initials: AMF
authors.lastname: Cerqueira
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro 20551-030, Brazil.
authors.forename: João R C
authors.initials: JRC
authors.lastname: Andrade
mesh_terms: D002460:Cell Line; D004705:Endocytosis; D004847:Epithelial Cells / Q000382:microbiology*; D006801:Humans; D050296:Microbial Viability; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D054323:Shiga-Toxigenic Escherichia coli / Q000502:physiology*; D013997:Time Factors; D014617:Vacuoles / Q000382:microbiology