Skip to main content
Fig. 3 | Journal of Biomedical Science

Fig. 3

From: Identification of human MHC-I HPV18 E6/E7-specific CD8 + T cell epitopes and generation of an HPV18 E6/E7-expressing adenosquamous carcinoma in HLA-A2 transgenic mice

Fig. 3

Characterization of HPV18 E6-specific CD8 + T cell immune response in wild type C57BL/6 mice vaccinated with therapeutic HPV vaccine. A Schema of the experiment. Briefly, female mice (5 per group) were vaccinated with 20 μg/mouse of pcDNA3-CRT/HPV18E6 DNA or pcDNA3-CRT/HPV18E6(delD70) vaccine on day 0 through intramuscular injection followed by electroporation. The mice were boosted once with the same dose and regime on day 7. Then mice were further boosted with 5 × 105 pfu/mouse of TA-HPV vaccinia vaccine through skin scarification on the tail on day 14. 12 days later, the spleens from the mice were pooled, and the splenocytes were prepared as described in the methods, stimulated with 15mer HPV18 E6 overlapping peptides that span the full-length of the HPV18 E6 protein at the presence of GolgiPlug overnight. 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 cells were acquired with FACSCalibur flow cytometer and data were analyzed with CellQuest Pro software. HPV18 E6 peptide-specific CD8 + T cells were determined as percent of IFN-γ + CD8 + T cells/total CD 8 + T cells. B HPV18 E6-specific CD8 + T cell responses in wild-type C57BL/6 mice after vaccination with DNA encoding CRT linked to wild-type HPV18 E6. C HPV18 E6-specific CD8 + T cell responses in wild-type C57BL/6 mice after vaccination with DNA encoding CRT linked to mutant HPV18 E6(delD70)

Back to article page