腺病毒表达Ribozyme在细胞内对HBV的免疫作用 |
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引用本文: | 李谨革,张嵘,周永兴,连建奇,王九平. 腺病毒表达Ribozyme在细胞内对HBV的免疫作用[J]. 解放军医学杂志, 2003, 28(3): 243-245 |
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作者姓名: | 李谨革 张嵘 周永兴 连建奇 王九平 |
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作者单位: | 710032,西安,第四军医大学唐都医院 |
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基金项目: | 国家自然科学基金资助课题(编号39570652) |
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摘 要: | 了解利用腺病毒腺体表达针对乙型肝炎病毒(HBV)的核酶在细胞内对HBV表达的抑制作用。将针对HBV的多位点核酶克隆入腺病毒载体中,利用包装细胞包装出含核酶及突变核酶的重组腺病毒,将腺病毒感染2.2.15细胞(含HBV),检测感染细胞中核酶及突变核酶的表达,利用ELISA,打点杂交及激光共聚光共聚焦等方法,检测核酶对细胞内乙型肝炎病毒表达的抑制作用。结果显示核酶及突变核酶的确被克隆入腺病毒载体,并经过包装后可形成包装腺病毒,感染2.2.15细胞后可表达出目的核酶,并发挥核酶的剪切作用,其对HBV表达的抑制率最高可达87.1%。证实包装腺病毒表达的多位点核酶对HBV的复制有明确的抑制作用,支持核酶的抗病毒效果,有希望成为HBV治疗的潜在方法之一。
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关 键 词: | 肝炎病毒 乙型 核酶类 基因疗法 |
修稿时间: | 2002-10-14 |
INHIBITION OF HBV BY RIBOZYME IN ADENOVIRUS EXPRESSION SYSTEM |
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Abstract: | To investigate the inhibition of HBV expression by multi-target ribozyme-expressing adenovirus vector in vivo, the ribozyme and muta-ribozyme were cloned into adenovirus vector pAd-CMV. By using the adenovirus expression system, the two plasmids were co-transfected into 293 cell line with adenovirus gene recombinant plasmids pJM17. The recombinant adenovirus were detected by plaque assay. Then the recombinant adenovirus infected the 2.2.15 cells. Using ELISA, dot-bolt hybridization and image analysis system, the inhibition of HBV expression was detected. The ribozyme and muta-ribozyme were cloned into the adenovirus vector pAd-CMV. The recombinant adenovirus could infect 2.2.15 cells and express the ribozyme. The ribozyme could decrease the expression of HBeAg by 87.1%. All these results strongly indicate that ribozyme can inhibit HBV expression in the cell, which represents a potential gene therapy for HBV infection. |
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Keywords: | hepatitis B virus ribozyme gene therapy |
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