Figure 8
Characterization of the E7-specific CD8+ T cell immune responses in iNOS and TNF-α knockout mice treated with HPV-16 E7 DNA vaccine in combination with DMXAA. A & B. Schematic diagram of the immunization regimen of the CRT/E7 DNA vaccine and DMXAA administered at different time points. 5-8 weeks old wild-type or iNOS deficient (middle panel), or TNF-α deficient (bottom panel) C57BL/6 mice were vaccinated with pcDNA3-CRT/E7 via gene gun delivery at day 0, and boosted once at day 7. The mice were treated with DMXAA (20 mg/kg) via i.p. injection either at day 0 (A) or day 3 (D) of the first vaccination. Splenocytes were harvested 7 days after last vaccination, and HPV-16 E7aa49-57-specific-CD8+ T cell responses were analyzed by intracellular IFN-γ staining using flow cytometry. B & C. Bar graphs depicting the number of E7-specific IFNγ+ CD8+ T cells per 3 × 105 splenocytes in WT, iNOS-/-mice (B) or TNF-α-/- mice (D) ± SEM following DNA vaccination and DMXAA treatment on D0. E & F. Bar graphs depicting the number of E7-specific IFNγ+ CD8+ T cells per 3 × 105 splenocytes in WT, iNOS-/-mice (E) or TNF-α-/- mice (F) ± SEM following DNA vaccination and DMXAA treatment on D3.The data shown here are from one representative experiment of two performed.