We have observed two patients who exhibited an exclusive increase of delta TCS1+ subset of gamma delta T cells in the peripheral blood after bone marrow transplantation (BMT). In one case with severe combined immunodeficiency (SCID) who received haploidentical BMT from his father, gamma delta T cells appeared only after thymus transplantation. However, his T cell-mediated immunity remained severely defective despite the generation of T cells of donor origin. In the other case with aplastic anemia, delta TCS1- gamma delta T cells began to increase in the peripheral blood later. This indicates that the thymus is necessary for the generation of gamma delta T cells and that the delta TCS1+ subset is dominant in the early stages of their ontogeny. delta TCS1+ T cell lines were established from both patients, and allo-reactivity was investigated. The cell line from the latter case reacted to recipient cells in a mixed lymphocyte reaction, but did not show cytotoxity to the allogeneic cells including recipient cells. The other cell line, from the former case, did not react to either donor or recipient cells. This indicates that an intact thymus is needed for gamma delta T cells to acquire allo-reactivity. Both cell lines showed MHC non-restricted cytotoxity against NK-sensitive target cells.