The aim of this study was to make unbiased quantitative estimates of the number of T8 cytotoxic cells and T4 helper cells and macrophages in the head and tail of rat epididymides, using newly developed design-based stereological methods in fertile control animals, and to compare these with results from vasectomized animals. The location of these cells within the intra- and interepithelial compartments of the epididymis was also studied. In control animals, both the T4 helper cells and macrophages and T8 cytotoxic cells were found in significantly higher numbers (P < 0.01) in the head interepithelial location than in the intraepithelial site. In addition, there were significantly more (P < 0.01) T8 cytotoxic cells in the tail interepithelial region. Within the head region of vasectomized rats, there was a significant increase in T8 cytotoxic cells in the intraepithelial location (P < 0.01), and a decrease in T4 helper cells and macrophages in the interepithelial region (P < 0.05). Serum samples from vasectomized animals, when compared with those from control animals, showed a significant increase in cytotoxic antisperm antibody. This suggests that, in response to the increased spermatozoal antigenic absorption that takes place in the epididymis after vasectomy, the immune system redistributes its T cell subsets to modify the immune tolerance that may occur.