Gamma-Glutamyltranspeptidase (GGT) catalyzes the first step in the extracellular hydrolysis of glutathione (GSH) and plays a critical role in GSH recycling; however, little is known about the impact of diminished GGT activity on immune function. We report here that GGT knockout (GGT(-/-)) mice have a 30 % decrease in splenic GSH and a 50 % reduction in thymus and spleen cellularity. The decreased cellularity was not selective for one population of cells, as each population was equivalently reduced. Following antigen challenge, GSH levels were reduced by 20-40 % in CD4+ and CD8+ T cells from GGT(-/-) mice when compared to T cells from wild-type mice. To test whether decreased GSH impairs immunity, we examined immune responsiveness following in vivo challenge with four different T cell-dependent stimuli. While there was no alteration in the antibody response to ovalbumin and sheep erythrocytes, cytotoxic T lymphocyte and alloantibody activity against P815 cells were decreased by 30 % and 65 %, respectively. Compared to wild-type littermates, anti-CD3-induced IL-2 and IL-6 production were also diminished in GGT(-/-) mice. These results demonstrate differential effects of decreased GSH on in vivo immune responsiveness to distinct stimuli, and suggest an important immunoregulatory role for GSH.