Shiga toxin kills epithelial cells isolated from distal but not proximal part of mouse colon.

Shiga toxins (Stxs) are major virulence factors produced by enterohemorrhagic Escherichia coli O157:H7 colonizing the human and cattle intestines. We previously demonstrated that recombinant binding subunits (Stx1B) bound to the mucosal epithelium of the distal but not that of the proximal part of the mouse colon. Here we developed a method for isolating colon epithelial cells from the proximal and distal parts separately. Enrichment of epithelial cells was confirmed by the expression of cytokeratin. There was no difference in the epithelial cell purity between the proximal and distal colon preparations. The isolated epithelial cells from the distal colon were found to display binding sites for recombinant Stx1B whereas those from the proximal colon were not. Taking advantage of this single cell isolation, we examined the effect of Stx1 holotoxin on the epithelial cells. Consistent with the expression of the binding sites, Stx1 induced apoptosis of the epithelial cells from the distal but not those from the proximal colon. The results provide direct evidence that mouse colon epithelial cells are susceptible to Stx1 toxicity corresponding to the expression of binding sites for toxins.