We have investigated the regulation of self tolerance in mice by examining lymphocyte reactivity in vitro against two subpopulations of autologous testicular cells: germ cells that were derived from the seminiferous tubules, and interstitial somatic cells. In the presence of germ cells, lymphocyte proliferation was strongly reduced. In contrast, somatic interstitial cells stimulated lymphocyte proliferation. In both cases, reactive lymphocytes were mostly T cells. Suppressor T cells activated by autologous germ cells were nonspecific and capable of inhibiting lymphocyte proliferation against autologous and allogeneic somatic testicular cells as well as against allogeneic spleen cells. Suppression was abrogated after treatment of the responder lymphocytes with anti-Ly-2.2 serum plus complement. Lymphocyte proliferation by autologous interstitial cells was considerably reduced, but not completely abolished, by complement-dependent lysis with anti-Thy-1.2 serum. This may indicate the participation in proliferation of a lymphoid cell population other than T cells.