Clinical evidence has revealed that NK cell function is critical to controlling infection with varicella zoster virus (VZV), the alphaherpesvirus responsible for varicella and herpes zoster. Despite this phenomenon, and the lymphotropic nature of VZV, understanding of NK cell interactions with VZV has remained surprisingly overlooked. We have demonstrated the first evidence of VZV infection of human peripheral blood NK cells, where strikingly 20–60% of cells were detected as antigen positive by flow cytometry. Importantly, NK cells were able to transmit infection to an epithelial cell monolayer, suggesting a possible role in the dissemination of virus. To determine whether a particular subset of NK cells was permissive to VZV infection we used FACS sorting of individual subsets, and high-dimensional computational interpretation of flow cytometry analysis of maturation markers. Through this we have identified preferential VZV infection of CD56dim CD57+ NKG2A– NK cells, correlating with a mature phenotype. However, while mature NK cells are reported to have high expression of Fc receptor CD16, VZV induced a potent loss of CD16 on the cell surface. Furthermore, we found by flow cytometry that VZV infection of CD57– NK cells remarkably drove expression of CD57. Of the VZV antigen positive cells, approximately 65% became CD57+ within 48 hours, and this effect was enhanced by concurrent stimulation with IL-2. Meanwhile the VZV antigen negative NK cells maintained low CD57 expression comparable to mock, under all conditions. Finally, whilst mature NK cells possess higher cytolytic potential than immature NK cells, we found that VZV-cultured NK cells no longer degranulated nor cytolytically killed target cells, when examined by flow cytometry for cell-surface CD107a and calcein-release cytotoxicity assay, respectively. Our findings identify not only a novel infection of mature NK cells, but provide evidence of a virus driving maturation in vitro, as well as interfering with cytolytic function.