Enterohemorrhagic Escherichia Coli Adherence Research Articles

Adherence of enterohemorrhagic Escherichia coli strains to a human colonic epithelial cell line (T84)

Enterohemorrhagic Escherichia coli (EHEC) produce Shiga-like toxins and attach to certain tissue culture cells. T84 cells are human colonic carcinoma cells. Unlike previously studied cell lines, T84 cells grown on collagen-coated surfaces polarize and produce tight junctions and desmosomes, forming a colonic epithelial cell layer in vitro. The purpose of this study was to examine the attachment of EHEC strains to the T84 cell line as a possibly more relevant in vitro model of EHEC adherence. Twelve EHEC strains were grown overnight in Penassay broth, suspended in minimal essential medium with and without 0.5% mannose, and incubated for 1 to 3 h with 5- to 7-day-old T84 cell monolayers grown on collagen-coated coverslips. The bacteria were removed, and attachment was quantitated microscopically. For both E. coli O157:H7 and other EHEC serotypes, there were marked differences in adherence between strains (range of 152 to 3 bacteria per oil immersion field). Mannose partially inhibited the adherence of some EHEC strains. Adherence to the T84 cells appeared to be related to the amount of pili present and not to the serotype. Electron micrographs showed that a highly adherent strain (strain 43-12) tended to form microcolonies in the area of tight junctions on the T84 cell monolayers. In addition, the attachment of these EHEC strains to T84 cells correlated with their ability to adhere to isolated rabbit colonocytes (r = 0.91, P = 0.00004; without mannose) (r = 0.60, P = 0.04; with mannose). These data show that there are EHEC strain-related differences in adherence which can be demonstrated in a human-derived colonic epithelial cell line (T84) and that these cells can be used to study EHEC adherence.

The Effect of Postnatal Age on the Adherence of Enterohemorrhagic Escherichia coli to Rabbit Intestinal Cells

Enterohemorrhagic Escherichia coli (EHEC) are associated with hemorrhagic colitis and hemolytic uremic syndrome. These illnesses are typically seen in young children, but are rare before 6 mo of age. The cause of this age restriction is unclear. Because bacterial adherence to intestinal mucosa is considered a critical initial event in pathogenesis, we studied the ontogeny of the adherence of EHEC (O157:H7 and other serotypes) isolated from children with diarrhea, hemorrhagic colitis, or hemolytic uremic syndrome. Adherence was quantitatively determined by incubating radiolabeled bacteria with viable rabbit intestinal cells, which were prepared by treating loops of distal ileum and proximal colon with EDTA, DTT, and citrate. Cells obtained from animals of different ages were studied simultaneously. The adherence of the various EHEC strains varied significantly. A non-O157:H7 E. coli strain 43-12 bound best (35 and 32 bacteria/cell to ileal and colonic cells, respectively) with 48-60% inhibition by D-mannose and alpha-methyl mannoside (p less than 0.01) and 20-28% inhibition by L-fucose (p less than 0.05), but no significant inhibition by other carbohydrates. Analysis of variance and polynomial regression showed that postnatal age significantly affected the adherence to ileal and colonic cells. Adherence during the 1st wk of life was 13-19% of that in the adult animal; it increased gradually, reaching the adult level at about 4 wk of age. Our study shows that postnatal age affects the adherence of EHEC to intestinal cells. These findings are compatible with postnatal development of gut receptors and may be relevant to the age-related risk of EHEC disease in children.