Hepatitis B virus inhibition in mice by lentiviral vector mediated short hairpin RNA.
Hepatitis B virus inhibition in mice by lentiviral vector mediated short hairpin RNA.
复制标题
慢病毒载体介导的短发夹RNA抑制小鼠乙型肝炎病毒
DOI：
10.1186/1471-230x-9-73
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发表时间：
2009-10-06
影响因子：
2.4
通讯作者：
Sun B
中科院分区：
文献类型：
--
作者：
Deng L;Li G;Xi L;Yin A;Gao Y;You W;Wang X;Sun B
研究背景慢性B肝炎病毒（HBV）感染是肝硬化和肝细胞癌的重要病因.目前治疗的主要挑战是目前药物的低疗效和耐药HBV突变的发生。针对病毒特异性基因的RNA干扰（RNAi）为开发新的抗HBV治疗提供了可能。最近的报道表明，基于HIV-1的慢病毒载体是有前途的基因递送载体，因为它们能够将转基因整合到非分裂细胞中。方法采用双抗体夹心酶联免疫吸附试验（ELISA）检测小鼠血清中B肝炎表面抗原（HBsAg）和B肝炎e抗原（HBeAg）的水平，并与正常对照组比较，用ELISA法检测小鼠血清中HBsAg和HBeAg的水平，用ELISA法检测小鼠血清中HBeAg的水平，用ELISA法检测小鼠血清中HBsAg和HBeAg的水平，用ELISA法检测小鼠血清中HBsAg和HBeAg的水平。免疫组化法检测小鼠肝组织中B型肝炎核心抗原（HBcAg）和HBsAg，分别采用荧光定量聚合酶链反应（RT-PCR）和实时荧光定量PCR检测HBV DNA和HBV mRNA。将HBV质粒与靶向HBV shRNA的慢病毒一起注射诱导RNAi应答。小鼠血清中分泌的HBsAg降低了89%，HBeAg也被显著抑制，肝组织中HBcAg或HBsAg的免疫组化检测也显示了显著降低。结论慢病毒介导的RNA干扰技术可有效抑制HBV在体内的复制，有望成为治疗HBV等病毒感染的一种有效手段。
BackgroundChronic hepatitis B virus (HBV) infection is an important cause of cirrhosis and hepatocellular carcinoma. The major challenges for current therapies are the low efficacy of current drugs and the occurrence of drug resistant HBV mutations. RNA interference (RNAi) of virus-specific genes offers the possibility of developing a new anti-HBV therapy. Recent reports have shown that lentiviral vectors based on HIV-1 are promising gene delivery vehicles due to their ability to integrate transgenes into non-dividing cells. Herein, a lentivirus-based RNAi system was developed to drive expression and delivery of HBV-specific short hairpin RNA (shRNA) in a mouse model for HBV replication.MethodsHepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the sera of the mice were analyzed by quantitative sandwich enzyme linked immunosorbent assay (ELISA) technique, hepatitis B core antigen (HBcAg) and HBsAg in the livers of the mice were detected by immunohistochemical assay, HBV DNA and HBV mRNA were measured by fluorogenic quantitative polymerase chain reaction (FQ-PCR) and quantitative real-time PCR respectively.ResultsCo-injection of HBV plasmids together with the lentivirus targeting HBV shRNA induced an RNAi response. Secreted HBsAg was reduced by 89% in mouse serum, and HBeAg was also significantly inhibited, immunohistochemical detection of HBcAg or HBsAg in the liver tissues also revealed substantial reduction. Lentiviral mediated shRNA caused a significant suppression in the levels of viral mRNA and DNA synthesis compared to the control group.ConclusionLentivirus-based RNAi can be used to suppress HBV replication in vivo, it might become a potential therapeutic strategy for treating HBV and other viral infections.