含有丙型肝炎病毒核心基因表达质粒的构建及其基因免疫

目的：研究丙型肝炎病毒（ＨＣＶ）核心（Ｃ）基因免疫诱生特异性免疫应答的可行性。方法：将ＨＣＶ Ｃ基因片段插入真核表达载体ｐｃＤＮＡ３质粒ＣＭＶ启动子的下游,构建真核表达载体ｐｃＤＮＡＨＣＶ－Ｃ,分别转染小鼠骨髓瘤细胞ＳＰ２／０和人肝癌细胞７７２１进行瞬时表达,用免疫荧光法和Ｗｅｓｔｅｒｎ－ｂｌｏｔ检测表达产物,将重组质粒注射,ＢＡＬＢ／ｃ（Ｈ－２＾ｄ）小鼠股四头肌,ＥＬＩＳＡ法检测血清中抗体产生水

Construction and gene immunization of recombinant expression plasmid of Hepatitis C virus core gene

Objective:To induce HCV specific humoral immune responses in mice using gene immunization with the core protein of HCV.Methods : The HCV core gene coding region was inserted into the plasmid pcDNA3, then the core gene expressed transiently with Lipofectamine coated in human hepatoblastoma 7721 and mouse myeloma SP2/0 cells.The product was detected by Western blot.The recombinant plasmid pcDNAHCV C was directly injected into the quadriceps muscle of BALB/c mice ,the specific antibody from the sera of immunized mice was measured by ELISA. Results: 7721 and SP2/0 cells were shown to express specific antigen, a 22 kD protein. HCV core antibody could be detected,and the titer of specific antibody rose as immune dose increased. Conclusion: The injection of pcDNAHCV C might elicit specific immune response in BALB/c mice, and be a promising candidate as antiviral agent and HCV genetic vaccine.