gamma delta T cells are known to localize preferentially in the epithelial regions and the hepatic sinusoids, and exhibit highly restricted V gene usage depending on their location. In the present study, gamma delta T cells in mice were further characterized in terms of their expression of the interleukin-2 receptor beta-chain (IL-2R beta), CD4 and CD8, and CD8 alpha and beta. This experiment was arranged to investigate whether gamma delta T cells have different properties depending on the organs and how gamma delta T cells are different from extrathymic alpha beta T cells, i.e. alpha beta T cells in the liver and intraepithelial lymphocytes in the intestine, in terms of the above phenotypes. Three-colour immunofluorescence tests using monoclonal antibodies revealed that gamma delta T cells can be classified into three groups: gamma delta T cells of the liver type are all IL-2R beta+, are comprised of double-negative (DN) CD8-CD4- and single-positive CD8+ (no CD4+) cells, and express CD8 alpha+ beta-; gamma delta T cells of the thymus type are a mixture of IL-2R beta+ and IL-2R beta-, are mainly DN, and express CD8 alpha+ beta+ if they carry CD8 antigens; and gamma delta T cells of the intestine type are also IL-2R beta+ or IL-2R beta-, are all CD8+, and express CD8 alpha+ beta-. gamma delta T cells in the spleen of normal mice are of the thymus type, while gamma delta T cells in the spleen of athymic nude mice seem to be of the liver type. All these properties of gamma delta T cells resemble those of extrathymic alpha beta T cells rather than regular alpha beta T cells of thymic origin. The present results reveal that gamma delta T cells and other extrathymic alpha beta T cells have many properties in common as primitive lymphocytes in phylogenetic development.