Figure 7. The nucleic acid vaccine exhibited significant immune and tumor-killing effects. (A) The titer of serum specific antibodies in mice was detected by ELISA at 4, 8, 12, 16 or 20 weeks. (B) IFN-γ levels in the culture supernatants of splenic lymph nodes co-cultured with stably transfected pIRES-neo3-HPV16 E6/E7 B16 cells were detected by ELISA. (C) The results of the ELISpot assay are pictured. (D) The ELISpot assay was used to detect the number of splenic lymphocytes secreting IFN-γ when the cells were co-cultured with stably transfected pIRES-neo3-HPV16 E6/E7 B16 cells. (E) The killing activities of splenic lymphocytes from pVAX1-IRES-GM-CSF-B7.1-HPV16 E6/E7 (HPV16) mice towards B16 cells stably transfected with HPV16 E6/E7 were tested with a cytotoxic lymphocyte killing test at lymphocyte:B16 cell ratios of 10:1, 20:1 and 40:1. *P < 0.05, **P < 0.01 compared to the NC group.