Skip to main content
. 2022 Apr 14;13:826164. doi: 10.3389/fimmu.2022.826164

Figure 5.

Figure 5

Generation and characterization of AdEpitopes (A) Schematic representation of oncolytic adenovirus delta 24 (Ad5/3-D24) constructs with modifications in E1, E3, and fiber region. Both unarmed (Ad_unarmed) and armed (Ad_CMV_Epitopes) bear a deletion of 24bp in the E1A gene. Additionally, AdEpitopes (Ad_CMV_Epitopes) contains an expression cassette under CMV promoter in E3 region. (B) Real-time PCR was performed in B16F1 infected with 5MOI of AdEpitopes, Ad5/3Δ24 or left untreated (not infected) and the fold gene expression is analyzed as 2-dCt.The data are represented ad bar blot and mean ± SEM. (C) IFN-γ ELISpot was performed on harvested splenocytes from mice treated with Ad5/3Δ24, AdEpitopes or PBS (Mock). The individual response to TRP2 (pink) and Ad5/3Δ24 (green) is reported as IFN-γ spot forming cells (SFC)/106 splenocytes. (D) IL-10 FluoroSpot evaluated the level of IL-10 released upon stimulation with TRP2 (pink) and Ad5/3Δ24 (green) in splenocytes harvested from mice immunized with Ad5/3Δ24, AdEpitopes or PBS (Mock). (E) The ratio IFN-γ/IL-10 spot forming cells is depicted and TRP2 (pink) and Ad5/3Δ24 (green). The data are shown as bar and dot plot for each technical replicate, and mean ± SEM. Significance was assessed with ordinary One-way ANOVA (ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001).