Figure 2

Gene expression associated with immune cell-mediated cytotoxicity is induced by the supernatant from mi-IL2. (A) Cytotoxicity was measured from co-cultures exposed to purified IL-2 or supernatant from mi-IL2 using Cytotox Green and the Incucyte imaging system. Three spheroids were analyzed for each condition. Supernatant from non-expressing bacteria were used as negative control (Neg.Ctrl). (B) Representative images of spheroid integrity after 3 days of co-culture. (C) RT-qPCR analysis of IL-2 response genes from tumor spheroid and PBMC co-culture. Gene expression was normalized to GAPDH expression. Mean values of relative gene expression and S.E.M. from three independent experiments are shown. qPCR primer sequences are listed in Table 3. (D) IFNγ levels from the tumor spheroid and PBMC co-cultures. Protein levels were measured by ELISA. Mean values of protein levels and S.E.M. from three independent experiments are shown. Significance determined with ANOVA and post-hoc analysis of comparison between groups using Tukey’s Honest Significant Difference test: (ns) p > 0.05, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.