Sensitivity in culture of epithelial cells from rhesus monkey kidney and human colon carcinoma to toxins A and B from Clostridium difficile.

The effect of toxins A and B from Clostridium difficile on human colon carcinoma cells (HT-29, epithelial), rhesus monkey kidney cells (MA-104, epithelial) and green monkey kidney cells (VERO, fibroblast) was studied. Both toxins caused rounding of HT-29 cells and rounding with projections remaining attached to the substrate in MA-104 and VERO cells; however, the sensitivity to each toxin varies considerably. Toxin A was detected in ng by VERO, pg by HT-29 and fractions of pg by MA-104 cells; for toxin B, pg were detected by VERO, ng by MA-104 and micrograms by HT-29 cells. HT-29 cells were grown with galactose to allow their differentiation to enterocytes, and their sensitivity to the toxins during the process was studied. At early stages, the sensitivity to both toxins was similar, and as the differentiation proceeded, the response to both toxins decreased continuously, and after 16 days no evident morphological effect was observed, even with micrograms amounts of either toxin. In contrast to all cell lines reported to date, HT-29 and MA-104 epithelial cells are exquisitely sensitive to toxin A and less responsive to toxin B. The rounding of HT-29 by these toxins depends on the degree of differentiation of the cell.