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Previous work in this laboratory showed that diethylstilbestrol was capable of suppressing induced furunculosis in rabbits. The present study indicates that the synthetic estrogenic hormone diethylstilbestrol which is used for acne, estrogen deficiency, cancer, and other disorders, can reduce the cytolytic action of staphylococcal alpha toxin. The cytotoxic action of purified alpha toxin for tissue cultures was evaluated by use of such parameters as total and viable cell counts, glucose, and protein determination, and cytopathic effects (CPE) in the presence and absence of steroids. To 3-day-old primary rabbit baby kidney tissue cultures, 1 to 5 mug of diethylstilbestrol per ml was added; growth of tissue cultures in Eagles medium was continued till the 6th day, and then one tissue cytopathic dose per milliliter of alpha toxin was added, and the subsequent fate of tissue cultures was assayed. Such cultures yielded higher total and viable cell counts, utilized more glucose, and contained more protein than the control cultures. In control cultures, CPE was observed on the 3rd hr after the addition of alpha toxin, and it was complete in 24 hr, whereas in tissue cultures treated with diethylstilbestrol, the CPE was significantly reduced. The data presented in this study made possible the availability of a suppressor of the cytolytic action of alpha toxin and might be useful in assaying the action of alpha toxin in an in vitro inexpensive test system.

The effect of the widely employed oral contraceptive steroid, Enovid, on the cytolytic action of the staphylococcal alpha toxin was investigated as an extension of previous studies in which it was shown that steroids were capable of suppressing induced staphylococcal infection in experimental animals. The cytotoxic action of alpha toxin for tissue cultures was evaluated by use of such parameters as total and viable cell counts, glucose and protein determination, and cytopathic effects in the presence and absence of Enovid. To 3-day-old primary rabbit baby kidney tissue cultures a mixture of 20 mug of norethynodrel per ml [17alpha-ethynyl-hydroxy-5(10)-estren-3-one] and 5 mug of mestranol per ml (17-ethynelestradiol-3-methyl ether) was added; growth of tissue cultures in Eagle medium was continued till the sixth day, and then one tissue cytopathic dose of alpha toxin per ml was added and the subsequent fate of tissue cultures was assayed. Such cultures yielded higher total and viable cell counts, utilized more glucose, and contained more protein than the control cultures. In control cultures, cytopathogenicity appeared on the third day after the addition of alpha toxin, and it was complete in 24 hr, whereas in tissue cultures treated with Enovid cytopathogenicity was significantly reduced. Thus the mixture of synthetic hormones known as Enovid, in pharmacological concentrations, was found capable of reducing the cytopathic action of alpha toxin, but only to a slightly lesser degree than such natural hormone as progesterone.