Pathogenesis of Shigella diarrhea. XIV. Analysis of Shiga toxin receptors on cloned HeLa cells.

Binding kinetics of Shiga toxin to HeLa CCL-2 cells and to cell lines cloned by limiting dilutions were determined. Lines with a wide range of sensitivity to Shiga toxin were obtained. Binding data, analyzed by a computer-based Scatchard model program, revealed two classes of binding sites, one of low affinity and high capacity and one of high affinity and low capacity. The number of high affinity, but not low affinity, sites present on the clones correlated with their sensitivity to toxin. Tunicamycin-treated CCL-2 cells became resistant to Shiga toxin in parallel with a reduction in the capacity of the high-affinity site. Cell content of Gb3, the glycolipid receptor for Shiga toxin, decreased as the sensitivity of the cells diminished. These data show that a minority of Shiga toxin binding sites of HeLa cells are involved in the cytotoxic response and suggest that Gb3 is the high-affinity functional cytotoxin receptor.