Jin Young-Hee, Hong Seung-Ho, Kim Kyongmin, Shin Ho Joon, Park Sun
Department of Microbiology, Ajou University School of Medicine, San 5 Wonchon-dong Youngtong-gu, Suwon 443-749, Korea.
Yonsei Med J. 2006 Oct 31;47(5):721-8. doi: 10.3349/ymj.2006.47.5.721.
Replication of the hepatitis B virus is suppressed by deficiency of the X protein. Although several molecules that block cellular targets of X protein reduce the production of hepatitis B virus progeny, the effect of a specific inhibitor of X protein on viral replication has not been investigated. To block X protein specifically, we adopted an intracellular expression approach using H7 single chain variable fragment (H7scFv), an antibody fragment against X protein. We previously demonstrated that cytoplasmic expression of H7scFv inhibits X protein-induced tumorigenicity and transactivation. In this study, intracellular H7scFv expression inhibits reporter gene transactivation but not viral replication determined by endogenous hepatitis B virus polymerase activity assay and real-time PCR. Our findings imply that intracellular expression of antibody fragment against X protein may not be an alternative therapeutic modality for inhibition of hepatitis B virus replication.
乙型肝炎病毒的复制会因X蛋白的缺乏而受到抑制。尽管有几种能阻断X蛋白细胞靶点的分子可减少乙型肝炎病毒子代的产生，但X蛋白特异性抑制剂对病毒复制的影响尚未得到研究。为了特异性阻断X蛋白，我们采用了一种细胞内表达方法，即使用H7单链可变片段（H7scFv），一种针对X蛋白的抗体片段。我们之前证明，H7scFv的细胞质表达可抑制X蛋白诱导的致瘤性和反式激活作用。在本研究中，通过内源性乙型肝炎病毒聚合酶活性测定和实时PCR确定，细胞内H7scFv的表达抑制了报告基因的反式激活，但并未抑制病毒复制。我们的研究结果表明，针对X蛋白的抗体片段的细胞内表达可能不是抑制乙型肝炎病毒复制的替代治疗方式。
