Immunity against Marek’s disease in chickens is mediated by both innate and adaptive host responses. This study aimed to determine the effects of Marek’s disease virus infection on distribution and frequency of γδ T cells in tissues, as well as their expression of cytokine genes. In the spleen of infected chickens, the number of γδ T cells increased by 10 and 21 days post-infection. Additionally, nearly 100% of the splenic γδ T cells in MDV-infected birds were CD8+ by day 21 post-infection. Conversely, the number of γδ T cells in the cecal tonsils of infected birds decreased. Splenic γδ T cells had up-regulated expression of interferon-γ early in infection followed by simultaneous gene expression of interleukin-10 during the later phases. In conclusion, these results suggest a potential role for γδ T cells in immunity to MDV and further elucidate the underlying immunological mechanisms that mediate immunity to MD.