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Human Papilloma Virus (Human papillomavirus, HPV) is a papovavirus Branch, a class of specific infection of the skin and mucous membranes of the double-stranded closed circular DNA virus, species-specific, and can not be grown in culture, the main infection human epidermal cells or mucosal epithelium, causing the site of infection lesions occur. Has isolated more than 100 HPV, is divided into low-risk and high-risk type. Low-risk type main 6,11,34,44-related mucosal epithelium of benign cause papilloma, condyloma, low-grade uterine intraepithelial neoplasia (CIN), high-risk 16,18,31 33,35,39 type, and cervical cancer is closely related to the organization of more than 90% of cervical cancer can be detected HPV, HPV16 proportion of the highest detection rate of more than 50%, so in vaccine research and development focused HPVl6 research , has a very important significance for the prevention and treatment of cervical cancer. HPV DNA is divided into three parts, early genes (E zone), late genes (L District), non-coding long control region (Long control region, LCR). E Zone 6 open reading frame (El, E2, E4, E5, E6, E7), where high-risk HPV E6 E7 protein encoded by the normal cells malignant transformation protein, and jointly promote viral replication, the infected cells persistent hyperplastic and tumor suppressor genes failure HPV E6 with the tumor suppressor gene p53 role it lost the DNA in the cell cycle error correction function, HPV E7 inhibition of pRb turn activation-induced genes that promote cell cycle continuing cycle bring the organization continues to hyperplasia, E6, E7 genes play an important role in cervical cancer sustainable development in. The development of a safe, effective, economical HPV vaccine, has a very important significance for the prevention and treatment of cervical cancer. This study successfully constructed of HPV16E6E7 gene recombinant plasmid pcDNA-E6E7, mice immunized with evaluation HPV16E6E7 gene vaccine immune effector. Method: digestion of recombinant plasmid pGEM-T-E6E7 E6E7 gene inserted into the eukaryotic expression vector pcDNA3.1 (-), to build HPV16E6E7 gene recombinant plasmid pcDNA-E6E7 transfection identified by restriction endonuclease and sequence detection COS-7 cells can be verified by IFA method HPV16E6E7 transient expression in eukaryotic cells in vitro. To build a successful recombinant plasmid, after a lot of preparation and purification, polyethylene amine (polyethylenimine, PEI) for the adjuvant parcel, intramuscular injection of 6-week-old BABL / c mice, and at the same time set the original vector pcDNA3.1 (-) and physiological saline as a control group, injection every two weeks once injected a total of three times in the last two weeks after immunization in mice fundus vein blood serum was separated, the IFA assay gene immunization in mice induced humoral and cell immune response. Results the build HPV16E6E7 recombinant plasmid pcDNA-E6E7 to transient expression in COS-7 cells, anti-E7 antibodies can be detected in the serum of mice immunized with the original vector pcDNA3.1 (-) and saline injection, not detected specific anti E7 antibody; generated by quantitative ELISA cytokine IL-2 and IFNγ secretion experimental group of IL-2 was significantly higher with the blank group, P LT; 0.01; experimental group of IFNγ higher than those in the control group and the blank group, P lt; 0.05 differences between groups were statistically significant. The experimental results show that the the gene vaccine build HPV16E6E7 transient expression analysis of the vaccine strain specific humoral immune response and cellular immune responses induced in mice immunized mice by IFA and ELISA quantitative detection in cells cultured in vitro. In this paper, the basic research of gene vaccine for HPV 16E6E7 HPV infection diseases vaccine experiment platform, looking for an effective method for the treatment of HPV, made a useful attempt for the HPV16 E6E7 gene vaccine development. Therefore, the research will provide the next step gene vaccine to treat HPV infection diseases preclinical research laid the foundation, at the same time, the results of this study will have potential applications in the control of HPV infection diseases.
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