Fig. 2

Comparison of HPV16 E7-specific CD8+ T cell responses induced by TA-CIN vaccination at different dose and vaccination route when combined with pNGVL4a-Sig/E7(detox)/HSP70 DNA vaccination. Five to eight weeks old female C57BL/6 mice (5 mice/group) were vaccinated with either 25 μg/mouse of pNGVL4a-Sig/E7(detox)/HSP70 DNA in 50 μl via intramuscular injection (leg muscle) or indicated dose of TA-CIN in 20 μl via either i.d. or i.m. injection. The mice were primed as indicated twice with 1-week interval. Seven days after last vaccination, PBMCs were prepared and stained with anti-mouse CD8 and HPV16 E7 tetramer. Splenocytes were prepared and stimulated with 1 μg/ml of HPV16 E7aa49–57 peptide at the presence of GolgiPlug (1 μl/ml) overnight at 37 °C. The cells were then stained with anti-mouse CD8 followed by intracellular IFN-γ. The data were acquired with FACSCalibur and analyzed with CellQuest. a Schematic illustration of the experiment. b and c Flow cytometry analysis of HPV16 E7-specific CD8+ T cells in peripheral blood. d and e Flow cytometry analysis of HPV16 E7-specific CD8+ T cells in spleen