In the past few years, cell-derived membrane vesicle-based tumor vaccines have been considered as valuable new tools for cancer immunotherapy. Despite promising results in cancer clinical trails, an improved method is urgently needed for high efficiency tumor vaccines for a broad spectrum of tumors. Here we developed a single membrane vesicle-based vaccine, which is active in repressing both melanoma (B16) and Lewis lung carcinoma (LLC) tumor growth. By using the intrinsic function of dendritic cells in the processing and presentation of antigens, we generated dendritic cell (DC)-derived membrane vesicles (DC-mv) bearing tumor antigens from both B16 and LLC cells. Vaccination with this DC-mv-based dual vaccine induced specific cytotoxic T lymphocytes (CTL)-dependent tumor rejection and suppressed the growth of both types of tumor xenografts in mice. In addition, induction of CTL by this vaccine resulted in cross-protection responses and consequently enabled significant enhanced anti-tumor effects, indicating the synergistic anti-tumor activity. Our study suggests that the DC-mv-based vaccine holds great potential as a highly effective, versatile, cell-free vaccine for inhibition of multiple types of tumor growth. (C) 2012 Elsevier Ltd. All rights reserved.