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Figure 3 | Journal of Translational Medicine

Figure 3

From: Combination immunotherapy and active-specific tumor cell vaccination augments anti-cancer immunity in a mouse model of gastric cancer

Figure 3

Tumor-specific IFN-γ release and cell-mediated cytotoxicity after vaccination with mGC8 cells and GM-CSF. T cells generated from TVDLN at day nine after vaccination were polyclonally activated and expanded as described in the Methods section and tested for tumor-specific IFN-γ release and cell-mediated cytotoxicity. In the cytokine release assay, T cells were either cultured alone, with an anti-CD3 antibody, with a syngeneic but unrelated tumor, MCA 310, with the related tumor cells 424GC or with mGC8 cells. Supernatants were harvested 18 h later for quantification of IFN-γ (and IL-5, not shown) by ELISA. (A) Vaccination with mGC8 with or without LP, GM-CSF, and IFA. Data are presented as the mean of two independent experiments in which co-cultures were performed in duplicate (± SE). IFN-γ secretion was significantly increased in the mGC8 GM-CSF/IFA group (p < 0.05) compared with the mGC8-, mGC8 IFA-, and LP mGC8 IFA-groups. LP, induction of lymphopenia followed by reconstitution. (B) Vaccination with mGC8 cells; TVDLN were additionally co-cultured with the TAg peptides T1 and T2. Means of duplicate measurements and SE are indicated (n = 4 for tumor cell lines and the non-stimulated control). (C) Cytotoxicity of TVDLN against mGC8 (black symbols) and MCA310 (open symbols) at declining effector-to-target cell ratio (E:T). Means of duplicate measurements (+/- AVEDEV) are shown. The experiment was repeated after restimulation of the LN cells with irradiated mGC8 tumor cells (10:1) followed by 5 days culture in CM supplemented with 60 IU/ml IL-2 revealing similar results (not shown). AVEDEV: average of the absolute deviations of the numbers above from their mean.

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