Translocation of Enterococcus faecalis strains across a monolayer of polarized human enterocyte-like T84 cells.

TitleTranslocation of Enterococcus faecalis strains across a monolayer of polarized human enterocyte-like T84 cells.
Publication TypeJournal Article
Year of Publication2004
AuthorsZeng, J, Teng, F, Weinstock, GM, Murray, BE
JournalJ Clin Microbiol
Volume42
Issue3
Pagination1149-54
Date Published2004 Mar
ISSN0095-1137
KeywordsBacterial Translocation, Cell Culture Techniques, Cell Line, Endocarditis, Enterococcus faecalis, Enterocytes, Escherichia coli, Feces, Genetic Complementation Test, Humans, Plasmids, Reference Values, Urine
Abstract

We used a two-chamber system to study transcytosis of Enterococcus faecalis across monolayers of human colon carcinoma-derived T84 cells, which show structural resemblance to the native intestine. Among 16 E. faecalis isolates from different sources, the well-characterized strain OG1RF and 8 other isolates (2 endocarditis isolates, 1 urine isolate, and all 5 fecal isolates) showed translocation in this assay, while 6 clinical isolates (3 endocarditis and 3 urine isolates), the recipient strain JH2-2, and the control, Escherichia coli DH5alpha, had no detectable translocation. Of two OG1RF mutants involving the previously studied epa (enterococcal polysaccharide antigen) gene cluster, known to be needed for virulence and resistance to killing by polymorphonuclear leukocytes, one epa mutant (TX5179) was unable to translocate, while TX5180, with an epa disruption farther downstream, showed a moderate decrease in translocation relative to that of the wild-type strain OG1RF (P < 0.01), indicating that the epa gene cluster is important for translocation across a T84 monolayer. This observation was confirmed by complementation of the epa mutant (TX5179) with epa genes and restoration of its translocation ability. In conclusion, we have demonstrated translocation of at least some strains of E. faecalis across T84 monolayers, although strains differ considerably in this ability, and we have demonstrated that epa mutations can cause marked changes in successful translocation. These results suggest that this model may be a useful in vitro system for studying the process of translocation from the intestinal tract.

DOI10.1128/jcm.42.3.1149-1154.2004
Alternate JournalJ. Clin. Microbiol.
PubMed ID15004067
PubMed Central IDPMC356884
Grant ListR37 AI047923 / AI / NIAID NIH HHS / United States
R37 AI47923 / AI / NIAID NIH HHS / United States