In previous studies, it has been shown that the granulocyte macrophage-colony stimulating factor ( GM- CSF) or interleukin-2 ( IL-2) surface modified MB49 bladder cancer stem cells ( MCSCs) vaccine could induce a specific antitumor immunity and against bladder cancer in mice model respectively. However, whether combined administration of GM- CSF and IL-2 could produce specific immune responses to cancer stem cells ( CSCs) was uncertain. MCSCs were established and characterized. GM- CSF and IL-2 MCSCs vaccines were prepared and bioactivity was evaluated. The therapeutic, protective, specific, and memorial immune response animal experiments were designed. Tumor-specific cytotoxic T lymphocytes assay, enzyme linked immunosorbent assay, flow cytometry assay were performed to indentify whether vaccine caused an antitumor immunity. Streptavidin ( SA)- GM- CSF and SA- IL-2 MCSCs vaccines were prepared successfully. Such vaccines inhibited the volume of tumor and prolonged the survival of the mice in animal experiments. The express of IgG or IFN-c, the portion of dendritic cells, CD8+ and CD4+ T cells were highest in the combined vaccines group than the SA- GM- CSF vaccine group, the SA- IL-2 vaccine group, the MCSCs group and the PBS group. The combined of GM- CSF and IL-2 vaccines could induce better antitumor immunity than a vaccine alone. [ABSTRACT FROM AUTHOR]