ABSTRACT
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.
