Therapeutic approach to hepatitis C using single-chain antibody and DNA gene transfer

使用单链抗体和 DNA 基因转移治疗丙型肝炎

• 批准号: 08557040
• 负责人: HAYASHI Norio
• 金额: $ 8.83万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08557040/
• 关键词: Single-chain antibody; Hepatitis C virus; Hepatitis B virus; Virus particle; Packaging; HBc; Replicative intermediate; Hepatoma cell line; C型肝炎; single-chain抗体; DNAワクチン; 遺伝子治療; HBc抗原; 単鎖状体; sFv

A cDNA for single-chain antibody (sFv) against hepatitis B virus core protein (HBc), 1C9, was isolated. We also cloned a cDNA for control sFv (γ9) that could not react HBc. HB611 cells that carry three copies of HBV-DNA and produce HBV particles were transfected with 1C9 or γ9 and we established the HB611 cells that stably express 1C9 or γ9, respectively. We found no difference in production of each HBV transcript and protein between 1C9-expressing HB611 and γ9-expressing ones. However, expression of HBV replicative intermediate was inhibited in 1C9-expressing HB611 compared with γ9-expressing ones. These results suggested that sFv against HBc might inhibit the packaging of pregenome RNA into HBc particles. Thus sFv against viral capsid protein was shown to be useful for inhibition of encapsidation of viral genome and sFv against hepatitis C virus core protein may be a candidate for therapeutic application of hepatitis C.

分离了针对乙型肝炎病毒核心蛋白 (HBc) 1C9 的单链抗体 (sFv) 的 cDNA，我们还克隆了对照 sFv (γ9) 的 cDNA，该 sFv 不能与携带 HBV 三个拷贝的 HB611 细胞发生反应。 -用1C9或γ9转染DNA并产生HBV颗粒，我们建立了分别稳定表达1C9或γ9的HB611细胞。我们发现表达 1C9 的 HB611 和表达 γ9 的 HBV 转录物和蛋白质的产量没有差异，但是，与表达 γ9 的 HB611 相比，表达 1C9 的 HB611 中 HBV 复制中间体的表达受到抑制。 HBc 可能会抑制前基因组 RNA 包装成 HBc 颗粒，因此针对病毒衣壳蛋白的 sFv 被证明可用于抑制 HBc。病毒基因组和针对丙型肝炎病毒核心蛋白的sFv的衣壳化可能是丙型肝炎治疗应用的候选者。

项目成果

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