rBCG-HPV16L1-E7疫苗的构建和免疫原性研究  

Construction of rBCG-HPV16L1-E7 and Study on Its Immunogenicity

作　　者：李翎 罗红敏[2] 王春英[2] 赵立民[2] 刘毅 

机构地区：[1]山东省职业卫生与职业病研究院,山东济南250062 [2]山东省肿瘤防治研究院,山东济南250117

出　　处：《预防医学论坛》2006年第3期257-262,共6页Preventive Medicine Tribune

基　　金：山东省科技攻关计划项目资助(NO.022100119)

摘　　要：[目的]构建以BCG为载体的rBCG-HPV16L1-E7疫苗,研究rBCG-HPV16L1-E7重组表达体嵌合乳头状瘤病毒样颗粒(VLPs)对小鼠免疫功能的影响,初步评价此疫苗对HPV相关肿瘤(宫颈癌)的预防作用。[方法]电穿孔法将pIJ702-HPV16L1-E7分泌型穿梭表达质粒导入BCG中表达,构成rBCG-HPV16L1-E7疫苗。Western blot检测E7蛋白的表达。采用rBCG-HPV16L1-E7疫苗皮下注射于C57BL/6小鼠,51Cr释放法体外分析免疫鼠的细胞毒性T淋巴细胞活性(cytotoxic T lymphocyte,CTL),ELISA法检测免疫鼠血清中E7特异性抗体和细胞因子的表达水平。[结果]Western blot法分析证实,该重组疫苗能特异地表达HPV16 E7蛋白。以rBCG-HPV16L1-E7免疫小鼠后,检测到的细胞因子IL-2、IFNγ及E7抗体的含量明显升高,诱导机体产生特异的抗体反应以及CTL反应。[结论]rBCG-HPV16L1-E7能增强小鼠的细胞和体液免疫反应,可作为HPV16预防性疫苗。[Objective]To construct a vaccine, rBCC-HPV16L1-E7 for HPV, and detect the specific CTI. effect of CD8^＋ T lymphocyte generated by the virus-like particles（VLPs） and evaluate the potency of it. [ Methods] BCG were transfected with the recombinant by electroporation assay. The transient expression of HPV16L1-E7 in the transfected BCG was assayed by western blotting, rBCG-HPV16 L1-E7 were vaccinated to mice, and spleen cells were isolated. The activities of cytotoxic T lymphocytes （CTL） isolated from immunized mice were tested in vitro with ^51Cr specific release assay. Cytokines and antibodies were detected by ELISA method. [Results]Western blotting verified that E7 gene could be expressed in BCG. Secretions of cytokines IL-2,IFNγ and antibodies increased by injection of rBCG-HPV16L1-E7 vaccinations. The CTLs and E7 specific antibody from immunized mice were efficaciously activated by rBCG-HPV16L1-E7 plasmid. [Conclusion] Cell and humoral immune reaction of mice could be strengthened by rBCG-HPV16L1-E7 vaccine.

关 键 词：人乳头状瘤病毒16型 疫苗 E7基因 L1基因 重组 BCG 电穿孔 病毒样颗粒 

分 类 号：R392[医药卫生—免疫学]