Figure 6
IFN-γ ELISPOT Analysis of CD8 T cells from Immunized Mice. CD8 T cells from immunized and control mice were purified by immunomagnetic sorting and co-incubated with the indicated APCs (protein-loaded PEC or tumor cells) for18 hours. A) 1 × 105 syngeneic PEC loaded with buffer (no protein), recombinant EGFP, or recombinant DEK were co-cultured with 1 × 105 CD8+ T cells from the following groups of mice: naïve (stripes), PC61 treated (gray), PC61 treated plus AGN2a-CD80/86 vaccinated (cultured in normal mouse serum (ms)), (white), and PC61 treated plus AGN2a-CD80/86 vaccinated (cultured in FBS), (black). B) 5 × 104 CD8 T cells purified from naïve mice, PC61-treated mice, or AGN2a-CD80/86+PC61 treated mice were co-cultured with 1 × 104 tumor cells: AGN2a (white), AGN2a/pcDNA3.1 (gray, vector only control), AGN2a/DEK (stripes), or cultured alone (black, T cells only control). Data is representative of three separate experiments. Each experiment consisted of T cells purified and pooled from 5 mice. Error bars represent the standard deviation calculated from triplicate ELISPOT wells.