Most adult peripheral blood gammadelta T cells express Vgamma9/Vdelta2-encoded TCR that recognize a restricted set of nonpeptidic phosphorylated compounds, referred to as phosphoantigens. They also express various MHC class I-specific inhibitory receptors (IR), in particular CD94/ NKG2-A heterodimers, which participate in the fine tuning of their TCR-mediated activation threshold. Most mature Vgamma9/Vdelta2 T cells express surface CD94 receptors, unlike cord blood or thymus-derived Vgamma9/Vdelta2 clones, thus suggesting a role for the microenvironment in IR expression. In the present study we show that most CD94- Vgamma9Vdelta2 PBL ex vivo express an intracellular pool of CD94/NKG2-A receptors that is translocated to the cell surface upon activation by phosphoantigens or IL-2. In stark contrast, intracellular CD94/NKG2-A complexes are undetectable in CD94- thymus or PBL-derived mature Vdelta2 T cell clones, and no surface induction is observed following phosphoantigen activation of T cell clones. Altogether these results provide new insights into the regulation of CD94/NKG2-A expression on T lymphocytes and suggest the existence of distinct mechanisms controlling in vivo and in vitro induction of IR on these cells.