丙型肝炎病毒核心基因免疫研究

目的研究丙型肝炎病毒（ＨＣＶ）核心（Ｃ）基因免疫用于预防和治疗ＨＣＶ感染的可行性和有效性．方法将ＨＣＶＣ基因片段插入真核表达载体ｐｃＤＮＡ３质粒ＣＭＶ启动子的下游，在证实其可以在小鼠骨髓瘤细胞ＳＰ２／０（Ｈ２ｄ）中表达之后，将重组质粒注射ＢＡＬＢ／ｃ（Ｈ２ｄ）小鼠股四头肌，ＥＬＩＳＡ检测血清中抗体产生水平；３ＨＴｄＲ掺入法测定免疫小鼠淋巴细胞ＨＣＶＣ抗原特异性增殖能力，４ｈ５１Ｃｒ释放法检测免疫小鼠细胞毒Ｔ细胞（ＣＴＬｓ）体外杀伤功能．结果免疫小鼠２０只，初次免疫２ｗｋ后，血清中均出现了ＨＣＶＣ抗体，且增加免疫剂量可提高抗体滴度；淋巴细胞增殖指数为６１０，明显高于对照组（Ｐ＜００１），ＣＴＬｓ体外特异性杀伤率为６３１％，也高于对照组（Ｐ＜００１）．结论ＨＣＶＣ基因免疫不仅可以诱导机体产生特异性的体液免疫，而且产生特异性的细胞免疫，它可能是防治ＨＣＶ的有效方法．

A study of gene immunization with recombinant expression plasmid of hepatitis C virus core antigen

AIM To study the efficacy and feasibility of DNA immunization with recombinant expression plasmid of hepatitis C virus core gene in prevention and treatment of HCV infection. METHODS The recombinant plasmid containing HCV C gene was constructed and expressed transiently by using lipofectamine in the SP2/0 cells. After appraisal and purification, these plasmid and control DNA were directly injected intramuscularly into Balb/c mice. The presence of HCV C specific antibody in the sera was checked by ELISA. Proliferative responses of spleen cells from experimental and control mice were determined by 3H thymidine incorporation, and CTL activity was measured by standard 4 h 51 Cr release assays by using SP2/0 cells transfected or uninfected plasmid pcDNAHCV C as targets. RESULTS All immunized mice developed anti HCV core antibodies, and this antibody was dose dependent. Lymphoproliferative responses were higher than from control mice ( P <0 05). Stimulation indices (SI) were 2 42-6 10. CTL activity assays showel that effector cells from mice immunized with recombinant HCV core DNA induced approximately 63 1% lysis of pcDNAHCV C transfected SP2/0 target cells. CONCLUSION The HCV core gene immunization is a promising method against HCV infection.