FIG. 2.

Mapping HPV-16 E5-specific CTL peptide epitope in C57BL/6 mice. Four- to six-week-old C57BL/6 mice were vaccinated and boosted 7 days later with rAd-E5. Two weeks after the first vaccination, splenocytes were harvested and stimulated with each group of peptides. Iintracellular cytokine staining with flow cytometry was then performed to determine the number of CD8⁺ IFN-γ⁺ double-positive cells. (A) Splenocytes from vaccinated mice were stimulated in vitro with each pool of peptides and stained for both CD8⁺ and IFN-γ⁺ antibodies. “Whole peptides” refers to all of the 16 synthesized peptides. The result of one representative assay from five similar independent experiments is shown. (B) Summary of the five independent experiments. The data represent the means and standard errors of five experiments. The y axis shows the percent positive cells, i.e., (the number of E5-specific CD8⁺ IFN-γ⁺ T cells/the total number of tested lymphocytes) × 100%.