Inhibition of protease-inhibitor-resistant hepatitis C virus replicons and infectious virus by intracellular intrabodies
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- 作者：Meital Gal-Tanamy ; Romy Zemel ; Larissa Bachmatov ; Rohit K. Jangra ; Assaf Shapira ; Rodrigo A. Villanueva ; MinKyung Yi ; Stanley M. Lemon ; Itai Benhar ; Ran Tur-Kaspa
- 关键词：Hepatitis C virus ; Intrabodies ; NS3 protease ; Inhibitors ; RNA replicons ; Single-chain antibodies
- 刊名：Antiviral Research
- 出版年：2010
- 出版时间：October 2010
- 年：2010
- 卷：88
- 期：1
- 页码：95-106
- 全文大小：1213 K
文摘
Hepatitis C virus (HCV) infection is a common cause of chronic liver disease and a serious threat to human health. The HCV NS3/4A serine protease is necessary for viral replication and innate immune evasion, and represents a well-validated target for specific antiviral therapy. We previously reported the isolation of single-chain antibodies (scFvs) that inhibit NS3/4A protease activity in vitro. Expressed intracellularly (intrabodies), these scFvs blocked NS3-mediated proliferation of NS3-transfected cells. Here we show that anti-NS3 scFvs suppress HCV RNA replication when expressed intracellularly in Huh7 hepatoma cells bearing either subgenomic or genome-length HCV RNA replicons. The expression of intrabodies directed against NS3 inhibited the autonomous amplification of HCV replicons resistant to small-molecule inhibitors of the NS3/4A protease, and replicons derived from different HCV genotypes. The combination of intrabodies and interferon-α had an additive inhibitory effect on RNA replication in the replicon model. Intrabody expression also inhibited production of infectious HCV in a cell culture system. The NS3 protease activity was inhibited by the intrabodies in NS3-expressing cells. In contrast, cell-free synthesis of HCV RNA by preformed replicase complexes was not inhibited by intrabodies, suggesting that the major mode of inhibition of viral replication is inhibition of NS3/4A protease activity and subsequent suppression of viral polyprotein processing.