Action of bis(betachloroethyl)sulphide (BCES) on human epidermis reconstituted in culture: Morphological alterations and biochemical depletion of glutathione.

Human keratinocyte cultures were treated with bis(betachloroethyl)sulphide (BCES), an alkylating and vesicant agent. At concentrations of 5 x 10(-4) to 5 x 10(-3)m, spontaneous detachment of the epithelium from the culture plate was observed, reproducing in vitro the cutaneous vesication observed in vivo. Progressive cellular alterations were shown with increasing concentrations of BCES (5 x 10(-5) to 5 x 10(-3)m). At low concentrations (5 x 10(-5)m), lesions of the nucleus, a significant target for BCES, were observed, along with lesions in the cytoplasmic organelles. An acute, dose-dependent depletion of cellular glutathione was observed, which occurred within 1 hr of treatment. Mechlorethamine, an analogue of BCES, induced at equivalent doses the same glutathione depletion and similar spontaneous detachment in vitro. We suggest that BCES, in addition to its genetic effects, acts by direct metabolic toxicity and induces glutathione depletion by direct conjugation. The lesions obtained in vitro reproduced those observed in vivo. Human keratinocyte cultures can be proposed as a good model for the study of the mechanisms of action of BCES.