Figure 7 Assay for tumor-specific, CTL activity and IFN-γ secretion

in immunized mice. (A), Splenic T cells from immunized mice were restimulated ex vivo by culturing with MMC-treated, MFC tumor cells. The restimulated T cells (effector cells) were incubated with target MFC or B16F10 cells for 20 h. Cytolytic activity (lysis) was determined. (B), Supernatants were collected for IFN-γ assay. All data are shown as means ± SD for 10 mice per group and are representative of three independent experiments. * P < 0.05. Finally, administration of Copanlisib in vivo DC-Ad-MAGE-1 was tested as a possible therapeutic benefit for distant, established visceral metastases. In this treatment model, the benefit of CCL3 and CCL20-recruited DCs as a tumor treatment was quantified by counting metastatic foci in selleck products pulmonary tumor-bearing mice. These were established by i.v. administration of 5 × 105 this website viable MFC tumor cells. Metastatic lung tumors were observed at day 3 after tumor cell implantation. Subsequently, tumor-bearing mice were treated with 1 × 106 DC-Ad-MAGE-1 cells in triplicate

at days 3, 7 and 11 after injection of tumor cells. As controls, mice were treated to the same regimen with either DC-Ad-LacZ, DC-MFC Ag, or untreated DCs. Visible lung metastases in these mice were counted in macrography at day 21 after tumor cell inoculation. Mice treated with DC-Ad-MAGE-1 showed a dramatic reduction in the number of lung metastatic foci. However, a decrease did not appear in mice receiving the control treatments 4-Aminobutyrate aminotransferase (Table 1). Table

1 Treatment of distant metastatic tumors with MAGE-1-modified DC vaccines Treatment Number of Lung metastases DC-Ad-MAGE-1 *31.38 ± 2.26 DC-Ad-LacZ 120.75 ± 2.71 DC-MFC Ag 77.25 ± 3.37 Untreated DC 124.38 ± 3.58 * P < 0.05, DC-Ad-MAGE-1 versus the other control groups. Discussion We have demonstrated that after injection of CCL3 and CCL20, F4/80-B220-CD11c+ DC precursors are quickly recruited into the peripheral blood. Furthermore, these CCL3 and CCL20-recruited DCs, when modified with tumor antigen gene MAGE-1, could induce not only an effective CTL response against gastric cancer cells ex vivo but also therapeutic, anti-tumor immunity in both subcutaneous tumor and pulmonary metastatic tumor models. Among many different immunotherapeutic strategies currently being evaluated, DC-based vaccination has attracted particular attention as a proven safe and potent therapy against tumors [14, 16]. Induction of tumor immunity can be initiated by effectors of innate immunity and can be further developed by cells of adaptive immunity, with DCs playing a central regulatory role.