HPV16E7E6抗原表位嵌合体DNA抗肿瘤疫苗的研究  

Anti-tumor chimeric DNA vaccines based on the expression of HPV-16 E7/E6 epitope genetically fused with the HSP 70N

作　　者：李轶杰[1] 陈开旭[1] 庄淑珍[1] 马正海[1] 张富春[1] 

机构地区：[1]新疆生物资源基因工程重点实验室新疆大学生命科学与技术学院,乌鲁木齐830046

出　　处：《中国免疫学杂志》2009年第7期606-609,616,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助(30860265);新疆生物资源基因工程重点实验室基金资助(No.XJDX0201-2005-02)

摘　　要：目的:探讨HPV16 E7、E6抗原表位与HSP70 N端重组DNA疫苗抗肿瘤活性。方法:以重叠PCR将HPV16 E7基因N端60个氨基酸的编码序列与E6基因的10个(48～57)氨基酸的编码序列及HSP70 N端序列进行融合。以pcDNA3.0为载体,构建真核表达载体pcD-E76HSP。重组质粒免疫C57BL/6小鼠后,通过淋巴细胞增殖实验和细胞毒性杀伤实验研究该疫苗激发的细胞免疫反应及反应强度;观察该疫苗对C57BL/6小鼠TC-1肿瘤细胞移植瘤的治疗效果。结果:重组DNA疫苗免疫C57BL/6小鼠后,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。结论:该疫苗能激发特异性细胞免疫反应,显著抑制HPV16转化的TC-1肿瘤细胞生长。Objeclive： DNA vaccines encoding the HPV16 E7 and E6 oncoproteins epitope genetically fused to the HSP70 N fragments were tested in mice for induction of T cell-mediated immunity and protection against tumor cell challenge. Melhods： The fusion gene （HPV16 E7aa1-60E-6 aa48-57-HSP70N-DNA, E76HSP） was produced with overlap PCR. The E76HSP-DNA gene was inserted into a mammalian expression vector,pcDNA 3.0,to construct the DNA vaccine candidate. Animals （C57BL/6 mice） were immunized with the vaccine,and cytotoxicity measurement and tumor protection assay were carried out to examine the immunological effects of the vaccine candidate. Results： Immunization with chimeric DNA vaccine induced HPV16-specific immune response and also conveyed protection from TC-1 induced tumor in vivo. A survival rate （60%） after 60 days of tumor challenge was observed. Conchlsion： The chimeric DNA vaccine can induce specific cellular immune responso in vitro and suppress HPV16 positive TC-1 tumor cell growth obviously in vivo.

关 键 词：人乳头状瘤病毒16型 抗原表位 热休克蛋白70 融合基因 DNA疫苗 

分 类 号：R737.33[医药卫生—肿瘤]