Inhibition of protease-inhibitor-resistant hepatitis C virus replicons and infectious virus by intracellular intrabodies

Meital Gal-Tanamy, Romy Zemel, Larissa Bachmatov, Rohit Jangra, Assaf Shapira, Rodrigo A. Villanueva, MinKyung Yi, Stanley M. Lemon, Itai Benhar, Ran Tur-Kaspa

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)95-106
Number of pages12
JournalAntiviral Research
Volume88
Issue number1
DOIs
StatePublished - Oct 2010
Externally publishedYes

Fingerprint

Replicon
Protease Inhibitors
Hepacivirus
Viruses
Peptide Hydrolases
RNA
Hepatovirus
Immune Evasion
RNA Replicase
Polyproteins
Single-Chain Antibodies
Serine Proteases
Virus Diseases
Virus Replication
Interferons
Antiviral Agents
Liver Diseases
Hepatocellular Carcinoma
Chronic Disease
Cell Culture Techniques

Keywords

  • Hepatitis C virus
  • Inhibitors
  • Intrabodies
  • NS3 protease
  • RNA replicons
  • Single-chain antibodies

ASJC Scopus subject areas

  • Virology
  • Pharmacology

Cite this

Inhibition of protease-inhibitor-resistant hepatitis C virus replicons and infectious virus by intracellular intrabodies. / Gal-Tanamy, Meital; Zemel, Romy; Bachmatov, Larissa; Jangra, Rohit; Shapira, Assaf; Villanueva, Rodrigo A.; Yi, MinKyung; Lemon, Stanley M.; Benhar, Itai; Tur-Kaspa, Ran.

In: Antiviral Research, Vol. 88, No. 1, 10.2010, p. 95-106.

Research output: Contribution to journalArticle

Gal-Tanamy, M, Zemel, R, Bachmatov, L, Jangra, R, Shapira, A, Villanueva, RA, Yi, M, Lemon, SM, Benhar, I & Tur-Kaspa, R 2010, 'Inhibition of protease-inhibitor-resistant hepatitis C virus replicons and infectious virus by intracellular intrabodies', Antiviral Research, vol. 88, no. 1, pp. 95-106. https://doi.org/10.1016/j.antiviral.2010.08.001
Gal-Tanamy, Meital ; Zemel, Romy ; Bachmatov, Larissa ; Jangra, Rohit ; Shapira, Assaf ; Villanueva, Rodrigo A. ; Yi, MinKyung ; Lemon, Stanley M. ; Benhar, Itai ; Tur-Kaspa, Ran. / Inhibition of protease-inhibitor-resistant hepatitis C virus replicons and infectious virus by intracellular intrabodies. In: Antiviral Research. 2010 ; Vol. 88, No. 1. pp. 95-106.
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