Intoxication of cultured human lung fibroblasts with Clostridium difficile toxin

The cytopathogenic effect of partially purified toxin from Clostridium difficile on cultured human lung fibroblasts was studied. Conditions for determination of 50% tissue culture dose were standardized. The cytopathogenic effect of the toxin was dependent on toxin concentration, exposure time, and density of the cells. Transfer of the cells to 0 degrees C did not inhibit binding of toxin to the fibroblast surface, but prevented the development of the cytopathogenic effect. Both binding of toxin and some intracellular step(s) were prevented by 2,4-dinitrophenol. These preventative effects were reversible. Before and concomitantly with the appearance of the cytopathogenic effect, the cellular uptake of uridine and of amino acids was markedly stimulated. Protein synthesis was depressed when 100% of the cells showed the cytopathogenic effect, but the synthesis of nucleic acids was inhibited only several hours later. The primary cellular target for the toxin is still unknown.