To investigate the role of gamma delta T cells in human heart transplantation, we searched for this T cell population in endomyocardial biopsies as well as in T cell lines and clones derived from graft-infiltrating lymphocytes. The number of gamma delta T cells in endomyocardial biopsies from transplanted patients (n = 55) was mostly low and did not differ significantly from nontransplanted patients (n = 21). Moreover, there was no association of gamma delta T cell distribution with rejection status or with time posttransplantation. Graft-derived T cell lines were established in the presence of autologous feeder cells and recombinant interleukin-2 to favor the growth of in vivo-activated T cells. Twenty T cell lines analyzed by flow cytometry showed low percentages of gamma delta T cells, and we were unable to obtain gamma delta T cell clones for functional studies. These results show that gamma delta T cells are poorly expressed on human heart allograft infiltrates and indicate that, when present, they are not activated in the graft. Our data suggest that gamma delta T cells do not have a major role in human heart rejection.