γδ T cells have been implicated in the immune response to microbial infections. A sub population of γδ T cells that express an unusual Vδ1 chain are thought to play a role in anti-viral immunity, although specific antiviral mechanisms remain largely unknown. To better understand how Vδ1+ γδ T cells respond to viral infection and whether this response shapes their phenotype, we mapped the cytokine profile, changes in frequency and phenotype of Vδ1+ γδ T cells in lung transplant patients that developed acute Human Cytomegalovirus (HCMV) infection. Peripheral Blood Mononucleocyte (PBMC) samples were collected from 27 patients who underwent a lung transplant and contracted or reactivated latent HCMV. Using multiparameter immunphenotyping of longitudinal samples before HCMV infection and post-HCMV, we were able to determine the cytolytic capacity, phenotype alteration and activation of Vδ1+ γδ T upon viral infection or reactivation. Notably, we found an increase in CD45RA+, CD27- cells (effector phenotype) Vδ1+ γδ T across all samples post-HCMV infection, indicating a shift in Vδ1+ γδ T cell cytokine expression as a result of HCMV exposure. Further, the Vδ1+ γδ T population in patients experiencing an active HCMV infection, displayed a greater capacity for cytotoxicity, with an increase in granzyme expression, particularly granzyme B, as well as upregulation of NKG2D and CD94. In summary, HCMV drives Vδ1+ γδ T cell activation and subsequent phenotypic changes and our results affirm the potential of Vδ1+ γδ T cells to respond to HCMV infection.