Fig. 2

Characterization of HPV16 E7-specific CD8 + T cell mediated immune response in HLA-A2 (AAD) transgenic mice vaccinated with CRT/E7(N53S) DNA vaccine. A Schema of the experimental regimen. Briefly, female HLA-A2 (AAD) transgenic mice (5 per group) were vaccinated with 10 μg/mouse of CRT/E7(N53S) DNA vaccine on day 0 through intramuscular injection followed by electroporation. One week later the mice were boosted twice with the same dose and regime at one week intervals. 6 days after the final booster, the splenocytes from the mice were prepared and stimulated with HPV16 E7 overlapping peptides that span the full-length of the HPV16 E7 protein, HPV16 E7aa11-20, E7aa49-57, E7aa82-90 or E7aa86-93 peptides. The cells were then stained with PE-conjugated anti-mouse CD8a. After permeabilization and fixation, the cells were stained with FITC-conjugated anti-mouse IFN-γ. The stained cells were analyzed by flow cytometry. B Bar graph to summarize the percent of activated (IFN-γ +) of HPV16 E7-specific CD8 + T cells out of the total CD8 + T cells in response to various peptide stimulation in HLA-A2 (AAD) transgenic mice after vaccination with CRT/E7(N53S) DNA