Macrophages, with potent cytotoxic and antigen-presenting activities, can be used in cancer treatment. The biological characteristics and antitumor effect of GM-CSF gene-modified and tumor antigen-pulsed macrophages were investigated. The high levels of GM-CSF could be detected in the supernatants of macrophages after gene transfer. The cytotoxicity and the expression of MHC class II molecules of the gene-modified macrophages increased significantly and the antigen-presenting ability was enhanced. The gene-modified macrophages were then pulsed with tumor antigen and used to treat the experimental pulmonary metastastic mice. The number of pulmonary metastases was reduced significantly and the cytotoxicity of the CTL induced from the splenocytes of the tumor-bearing mice also increased. The results demonstrated that adenovirus-mediated GM-CSF gene transfer can activate macrophages to some extent and GM-CSF gene-modified, antigen-pulsed macrophages may be a new type of effective effector cells in the immunogene therapy of cancer.