This study was aimed to investigate the effects of tumor antigen-loaded dendritic cells (DC) stimulating the specific cytotoxic T lymphocytes (CTL) on Jurkat cells in vitro. Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation from normal human heparinized blood, the adherent monocytes were cultured with granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4), alpha tumor necrosis factor (TNF-alpha) and sCD40L, DCs were co-cultured with frozen-thawed antigen of Jurkat cells or WT1 peptides, and then T cells were triggered into specific CTL. The results showed that most suspended cells exhibited distinctive morphological features of DC which expressed CD40 (96%), CD86 (97%), CD80 (77%), CD1a (69%), and gained the powerful capacity to stimulate proliferation of allogeneic lymphocytes. Under the effector: target ratio of 20:1, CTLs derived from cultures with DC and frozen-thawed antigen of Jurkat cells showed 91.1% cytotoxicity against Jurkat cells, CTL derived from cultures with DC and WT1 peptides showed 87.5% cytotoxicity against Jurkat cells, cytotoxicity of CTL derived from cultures with unloaded DC against Jurkat cells was 42.1% and cytotoxicity of monocytes was 22.7%. Cytotoxicity of CTL derived from culture with frozen-thawed antigen or WT1 peptides loaded DC was stronger than that in control groups (P < 0.01). It is concluded that the tumor antigen-pulsed DC can induce efficient and specific anti-tumor immunity, may play a great role in clinical therapy for leukemia.