Human long-term bone marrow stromal cultures (LTBMC) have been transduced to express the immunostimulatory cytokine interleukin-2 (IL-2). LTBMC were established from normal bone marrow and characterized by gross morphology, immunohistochemistry, immunophenotyping, and immunofluorescent staining. A stromal marrow cell (SMC) population expressing the markers CD68, CD29, CD13, and CD54 was identified. These cells were successfully transduced with an adenoviral vector containing the human IL-2 gene. Production of IL-2 was demonstrated for at least 13 days post-transduction and was maximal on day 10 at 19,350 pg (or 38 IU) of IL-2/10(6) cells/24 h. IL-2 expressed by the transduced SMC was demonstrated to be biologically active, resulting in the proliferation of autologous T lymphocytes in a mixed lymphocyte reaction. Autologous human LTBMC can be obtained and manipulated with relative ease and may be a useful vehicle for sustained local cytokine delivery.