Invasiveness as a putative additional virulence mechanism of some atypical Enteropathogenic Escherichia coli strains with different uncommon intimin types

BMC Microbiol. 2009 Jul 21:9:146. doi: 10.1186/1471-2180-9-146.

Abstract

Background: Enteropathogenic Escherichia coli (EPEC) produce attaching/effacing (A/E) lesions on eukaryotic cells mediated by the outer membrane adhesin intimin. EPEC are sub-grouped into typical (tEPEC) and atypical (aEPEC). We have recently demonstrated that aEPEC strain 1551-2 (serotype O non-typable, non-motile) invades HeLa cells by a process dependent on the expression of intimin sub-type omicron. In this study, we evaluated whether aEPEC strains expressing other intimin sub-types are also invasive using the quantitative gentamicin protection assay. We also evaluated whether aEPEC invade differentiated intestinal T84 cells.

Results: Five of six strains invaded HeLa and T84 cells in a range of 13.3%-20.9% and 5.8%-17.8%, respectively, of the total cell-associated bacteria. The strains studied were significantly more invasive than prototype tEPEC strain E2348/69 (1.4% and 0.5% in HeLa and T84 cells, respectively). Invasiveness was confirmed by transmission electron microscopy. We also showed that invasion of HeLa cells by aEPEC 1551-2 depended on actin filaments, but not on microtubules. In addition, disruption of tight junctions enhanced its invasion efficiency in T84 cells, suggesting preferential invasion via a non-differentiated surface.

Conclusion: Some aEPEC strains may invade intestinal cells in vitro with varying efficiencies and independently of the intimin sub-type.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Adhesins, Bacterial / genetics*
  • Enteropathogenic Escherichia coli / classification
  • Enteropathogenic Escherichia coli / genetics*
  • Enteropathogenic Escherichia coli / pathogenicity
  • Escherichia coli Proteins / genetics*
  • Genes, Bacterial
  • HeLa Cells
  • Humans
  • Microscopy, Electron, Transmission
  • Serotyping
  • Tight Junctions / metabolism
  • Virulence / genetics*

Substances

  • Adhesins, Bacterial
  • Escherichia coli Proteins
  • eaeA protein, E coli