Effect of six virustatic nucleoside analogues on the development of fetal rat thymus in organ culture

Abstract

The effects of the virustatic agents zidovudine (azidothymidine, AZT) 2′3′-dideoxycytidine (ddC), 2′3′-dideoxyinosine (ddI), acyclovir (ACV), ganciclovir (GCV), and vidarabine phosphate (VP) on the in vitro development of thymic lobes of 17-day-old rat fetuses were tested in an organ culture system. The virustatics were added to the medium for a culture period of 7 days. All nucleoside analogues inhibited the proliferation and differentiation of lymphatic cells. However, differences were observable with respect to the potency of the six drugs to interfere with thymic development. Compared to untreated controls, reduction in the number of thymocytes was significant at concentrations of 30 μM AZT and ddI. In the case of ACV, GCV, VP, and ddC concentrations as low as 10 μM were sufficient to cause a significant reduction, ddC being the most potent derivate. Increasing concentrations of the nucleoside analogues led to a dose-dependent further inhibition of cell proliferation. At a concentration of 30 μM flow cytometry revealed a decrease in the relative number of double positive CD4⁺ CD8⁺ and single positive CD4⁺ CD8⁻ cells but an increase in the relative number of CD4-CD8⁺ cells. At the same concentration the expression of the CD5 antigen was reduced by the antimetabolites, indicating that maturation of the thymocytes was inhibited. Distribution of the forward light scatter, a cell size-related parameter, showed that the formation of small thymocytes was reduced by the nucleoside analogues. Light and electron microscopic investigations indicated cytotoxic effects of the drugs on the thymocytes, whereas the epithelium was only slightly affected.