预防医学论坛2006年5月第12卷第3期 Prev Med .doc

预防医学论坛2006年5月第12卷第3期 Prev Med Trib Vol.12,No.3,May. 2006 预防医学论坛 06年5月第12卷第3期 Med Trib Vol.12,No.3,May. 2006 【论 著】 rBCGHPV16L1E7疫苗的构建和免疫原性研究* 李翎1，罗红敏2，王春英2，赵立民2，刘毅1 （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） 中图分类号：R392 文献标识码：A 文章编号：1672-9153(2006)03-0257-06 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， 51Cr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。 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 Construction of rBCGHPV16L1E7 and Study on Its Immunogenicity／〖WTBX〗LI Ling, LUO Hongmin,WANG Chunying,et al(Shandong Academy of Occupational Health and Occupational Medicine,Jinan,250062，Shandong，China) Abstract:［Objective］To construct a vaccine, rBCGHPV16L1E7 for HPV, and detect the specific CTL 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 51Cr 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］