Thymic regression and apoptosis in the rat after treatment with the Leydig cell cytotoxin ethylene dimethanesulphonate (EDS).

Ethylene dimethanesulphonate (EDS) is an alkylating agent which is widely assumed to specifically kill Leydig cells leaving other biological systems intact. However, after EDS treatment of the male rat the thymus reversibly involutes and the gonadal regional lymph nodes are activated. In the present experiments we have demonstrated that EDS has a direct action upon the thymus both in vivo and in vitro. EDS treatment of the intact and castrated male rat and the intact female rat caused regression of the thymus by up to 50% 3 days later. Total cellularity decreased while the proliferative index increased suggesting a compensatory mechanism. Thymocytes were exposed to EDS in vitro and the response compared to the glucocorticoid methylprednisolone (P), a well characterised thymic apoptotic stimulant. EDS and P increased apoptosis in the thymocyte as characterised by the appearance of cells containing nuclei with apoptotic morphology and with DNA fragmentation visualised by a characteristic ladder after agarose gel electrophoresis. The effects of both EDS and P were time and dose dependent but, in contrast to the effects in Leydig cells, P was the most effective apoptotic stimulus (for instance 100%-P compared to 30%-EDS or 7% control/DMSO after 24 h incubation). The immunological responses of the gonadal lymph nodes were not associated with testicular regression as it was seen in the castrated rat but may be related to a direct action upon the epididymis. In conclusion, tissue specificity of the Leydig cell cytotoxin needs to be extended to the thymus and epididymis. The mode of cell death in Leydig cells and thymocytes after both glucocorticoids and EDS is apoptosis which suggests that they possess some common mechanism(s) which is responsible for the toxicity of these diverse compounds.