Immunization of dendritic cells (DC) pulsed with tumor antigen can activate tumor-specific cytotoxic T lymphocytes (CTL) which are responsible for protection from tumor challenge and regression of established metastatic tumor. It has been hypothesized that tumor lysate contains factors that may modulate DC maturation. In this study, we examined whether the uptake of tumor lysate (MCA-102 fibrosarcoma) could modulate DC phenotypes in vitro and whether the administration in vivo of tumor lysate-pulsed DC (TP-DC) could elicit efficient tumor specific immune responses followed by a regression of established tumor burdens. It was investigated the uptake of tumor lysate by DC by means of flow cytometry and fluorescent microscope. Murine bone marrow-derived DC efficiently phagocytosed tumor lysate and after the uptake, the phenotype of TP-DC was surprisingly comparable to unpulsed-DC (UP-DC), exhibiting lower levels of CD80 (<51%), CD86 (<43%), and MHC class II (<59%). Also, TP-DC did not enhance secretion of IL-12p70 (UP- vs. TP; 54.5+/-6.4 vs. 50.5+/-4.8 pg/ml, respectively), contrary to those activated with LPS (113.6+/-16.8 pg/ml). However, TP-DC vaccination in vivo increased the IFN-gamma production from splenocytes higher than that of UP-DC (TP- vs. UP-DC; 41029+/-1523 vs. 4752+/-590 pg/ml, respectively). Furthermore, the administration of TP-DC enhanced specific T cell responses against MCA-102 fibrosarcoma. These results demonstrate that augmentation of DC phenotype and function in vitro is not necessarily a prerequisite for TP-DC vaccination to successfully promote anti-tumor immunity in vivo.