Figure 3.

Analysis of HPV16/E7-specific systemic and local immune responses produced by intramuscular vaccination at different sites in TC-1 tumor-bearing mice. (A-B) Female C57BL/6 mice (five to eight weeks old, five/group) were injected with 2 × 10⁴ TC-1/luc cells at the intravaginal cavity on day 0. The mice were vaccinated with or without 20 μg/mouse of pNGVL4a-CRT/E7(detox) DNA vaccine via IM injection, followed by electroporation at different sites on day 3, and subsequently boosted again five days later. Peripheral blood lymphocytes were prepared one week after the final vaccination. The cells were stained with anti-mouse CD8 antibody, H2-D^b/E7 tetramer, and 7-AAD. A. Representative flow cytometry images of E7-specific CD8⁺ T cells in peripheral blood. B. Summary of E7-specific CD8⁺ T cells in peripheral blood. (C-D) Female C57BL/6 mice (five to eight weeks old, five/group) were injected with 5 × 10⁴ TC-1/luc cells at the intravaginal cavity on day 0. The mice were vaccinated with 20 μg/mouse of pNGVL4a-CRT/E7(detox) DNA vaccine via IM injection, followed by electroporation at different sites on day 4 and boosted once four days later. LNs were harvested seven days after the final vaccination and stained for HPV16/E7-specific CD8⁺ T cells. C. Representative flow cytometry images of E7-specific CD8⁺ T cells in LNs. D. Summary of E7-specific CD8⁺ T cells in LNs. (E-F) one week after the final vaccination, tumor tissues from mice described in subfigures A-B were harvested, and the tumor infiltrating lymphocytes were prepared and stained with anti-mouse CD8 antibody, H2-D^b/E7 tetramer, and 7-AAD. E. Representative flow cytometry images of E7-specific CD8⁺ T cells in TILs. F. Summary of E7-specific CD8⁺ T cells in TILs. Data are presented as mean ± SD. (* p < 0.05, ** p < 0.01, n.s. not significant).