小鼠及家兔对丙型肝炎病毒多表位DNA疫苗免疫应答的研究  

Immunogenicity of a multiple epitope antigen gene of hepatitis C virus in mice and rabbits

作　　者：黄建生[1] 陈丽珊[1] 雷呈祥[2] 解咏梅[1] 张潜[1] 沈先荣[2] 贾福星[2] 张丽芸 陈立茵 郭明秋 任大明[1] 

机构地区：[1]复旦大学生命科学院 [2]海军医学研究所,上海200433 [3]第一军医大学抗衰老研究室

出　　处：《细胞与分子免疫学杂志》2001年第1期52-54,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助!No.39780002

摘　　要：目的探讨ＨＣＶ复合多表位ＤＮＡ疫苗的可行性。方法把ＨＣＶ多表位抗原基因ＰＣＸ克隆到真核表达载体ｐＲＥＰ９（ＲＳＶ启动子）及ｐＣＤＮＡ３（ＣＭＶ启动子）中，构建真核表达载体ｐＲＥＰ９／ＰＣＸ及ｐｃＤＮＡ３／ＰＣＸ。将其肌肉注射免疫小鼠及家兔，检测特异性体液免疫和细胞免疫的水平，并观察免疫小鼠的安全性。结果质粒ｐＲＥＰ９／ＰＣＸ及ｐｃＤＮＡ３／ＰＣＸ于免疫后第 ６ｗｋ和第 １０ ｗｋ时，开始可检测到抗 ＧＺ－ＰＣＸ ＩｇＧ，随后抗体滴度逐渐升高，但水平较低，持续时间较短。ｐｃＤＮＡ３／ＰＣＸ肌肉注射免疫家兔后，于第８ｗｋ开始出现特异性抗体，至 ３个月后滴度升至 １： ３ ２００，随后也开始下降。免疫小鼠及家兔可诱发针对ＧＺ－ＰＣＸ融合蛋白的迟发型超敏反应，并刺激淋巴细胞转化。免疫后小鼠的体重正常，肝脾脏未见明显肿大，具有良好的安全性。结论ＨＣＶ多表位基因可诱发特异性免疫应答且安全性好，为ＨＣＶ疫苗的研究提供了一定的理论及实验依据。Aim To explore the possibility of the multiple epitope DNA vaccines of hepatitis C virus (HCV). Methods A synthetic multiple epitope antigen gene PCX of HCV was cloned into vector pREP9(RSV promoter) and pcDNA3 (CMV promoter) to construct eukaryotic expression vectors pREP9/PCX and pcDNA3/PCX, then they were used to immunize mice and rabbits, the titer of specific humoral and cellular responses were detected and their safety were observed. Results In mice, specific anti-GZ-PCX antibody(IgG) was lower than 1: 1 000 and did not persist well. In rabbits, the highest titer of anti-GZ-PCX IgG reached at 1: 3 200 and remained for about one month. Delayed type hypersensitivity reactions (DTH)and proliferation response of peripheral lymphocytes were induced by GZ-PCX antigen. Body weights of immunized mice were normal and no obvious toxic reaction was observed. Conclusion The multiple epitope antigen gene of HCV could induce specific immune responses without obvious toxicity and it might be able to serve as an effective HCV vaccine candidate.

关 键 词：丙型肝炎病毒 多表位抗原 基因免疫 免疫应答 

分 类 号：R512.630.3[医药卫生—内科学]