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γδ T cells are thought to mediate immune responses at epithelial surfaces. We have quantified and characterized hepatic and peripheral blood γδ T cells from 11 normal and 13 unresolved tumor-bearing human liver specimens. γδ T cells are enriched in normal liver (6.6% of T cells) relative to matched blood (0.9%; P = 0.008). The majority express CD4⁻CD8⁻ phenotypes and many express CD56 and/or CD161. In vitro, hepatic γδ T cells can be induced to kill tumor cell lines and release interferon-γ, tumor necrosis factor-α, interleukin-2 and interleukin-4. Analysis of Vγ and Vδ chain usage indicated that Vδ3⁺ cells are expanded in normal livers (21.2% of γδ T cells) compared to blood (0.5%; P = 0.001). Tumor-bearing livers had significant expansions and depletions of γδ T cell subsets but normal cytolytic activity. This study identifies novel populations of liver T cells that may play a role in immunity against tumors.