利用乳酸杆菌表达系统制备HPV16粘膜疫苗

项目名称： 利用乳酸杆菌表达系统制备HPV16粘膜疫苗

项目编号： No.30860265

项目类型： 地区科学基金项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 李轶杰

作者单位： 新疆大学

项目金额： 22万元

中文摘要： 本研究利用预测得的新疆株HPV16型特异的抗原表位，进行重组克隆，经DNA疫苗免疫C57BL/6小鼠后，流式细胞仪检测显示细胞因子表达以MHC-I型为主，小鼠脾淋巴细胞体外增殖明显，并可诱导产生针对TC-1肿瘤细胞的特异性CTL反应；体内抑瘤试验显示该疫苗对HPV16病毒转化的TC-1细胞小鼠移植瘤的生长有抑制作用。研究也分别证实3拷贝的鼠C3d分子与热激活蛋白70能够促进疫苗的激发效果，改进与优化了乳酸菌质粒的提取方法，为后续工作的顺利进行提供技术保障。研究将HPV16点突变的E7基因与鼠IL-12分别构建穿梭载体后转化乳酸菌，以此疫苗滴鼻免疫C57BL/6小鼠后，该乳酸菌疫苗能激发特异性细胞免疫反应，抑制HPV16相关的TC-1肿瘤细胞生长。结果显示该乳酸菌疫苗有望成为HPV16性相关肿瘤的预防与治疗型疫苗。

中文关键词： 人乳头瘤病毒；乳酸菌；疫苗；免疫；抗肿瘤

英文摘要： Putative cytotoxic T-lymphocyte (CTLs) epitopes of HPV16 cloned from HPV Xinjiang strain was inserted into different vectors to construct chimeric DNA vaccine and to immunize C57BL/6 mice. Flow cytometry analysis showed that the expressed intracellular cytokine was primary MHC-I type. Immunization of with chimeric DNA vaccine induced HPV16-specific immune response and also conveyed protection against TC-1 induced tumor in vivo. Surprisingly, chimeric DNA vaccines exclusively targeted CD8+ T cells; immunological and antitumor effects were completely CD4-independent. These results indicated that fusion of HSP70 or C3d3 to an antigen gene may greatly enhance the potency of DNA vaccines via CD8-dependent pathways. A simple, rapid and safe method was developed for extracting plasmid DNA from L.lactis. Point mutated HPV16 E7 gene and mouse IL-12 were used to construct the shuttle vector，and transformed into lactic acid bacteria. The lactic acid bacteria vaccine is able to stimulate specific cellular immune response, as vaccine intranasal immunization female C57BL/6 mice, and inhibit HPV16-related TC-1 tumor cell grow. Results demonstrated that the lactic acid bacteria including HPV16-E7m-DNA compound might become a candidate for HPV16 precautionary and immunotherapy.

英文关键词： Human papillomaviruses; Lactic acid bacteria; Vaccine; Immunization; Antitumor