Vγ9Vδ2 T cells can recognize various molecules associated with cellular stress or transformation, providing a unique avenue for the treatment of cancers or infectious diseases. Despite unique potential of these cells, the clinical efficacy of Vγ9Vδ2 T cell-based immunotherapeutics has to date been limited. Our work has focused on both in vitro and in vivo studies of Vγ9Vδ2 T cell function and trafficking. We have defined promising culture conditions that could improve Vγ9Vδ2 T-cell-based immunotherapies and shown in pre-clinical animal models that the tissue distribution and phenotype of in vivo pharmacologically expanded Vγ9Vδ2 T cells can be altered based on the antigen administration route. This work furthers our understanding of how these cells might recognize and target transformed or infected cells, implications for tissue trafficking and the clinical efficacy of Vγ9Vδ2 T cell immunotherapeutics.