Inhibition of hepatitis C virus NS3-mediated cell transformation by recombinant intracellular antibodies

Hepatitis C virus (HCV) infection is a major worldwide health problem, causing chronic hepatitis, liver cirrhosis and primary liver cancer. In addition to its role in the viral polyprotein-processing, the viral NS3 serine protease has been implicated in interactions with various cell constituents re...

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Bibliographic Details
Published in:Journal of hepatology 2004-06, Vol.40 (6), p.1000-1007
Main Authors: Zemel, Romy, Berdichevsky, Yevgeny, Bachmatov, Larisa, Benhar, Itai, Tur-Kaspa, Ran
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Biological and medical sciences
Cell Transformation, Viral
Cells, Cultured
Cloning, Molecular
Gastroenterology. Liver. Pancreas. Abdomen
Hepacivirus - drug effects
Hepacivirus - genetics
Hepacivirus - physiology
Hepatitis C virus
Human viral diseases
Infectious diseases
Intrabodies
Liver diseases
Medical sciences
Molecular Sequence Data
Plasmids
Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Serine protease
Tumorigenicity
Viral diseases
Viral hepatitis
Viral Nonstructural Proteins - chemistry
Viral Nonstructural Proteins - genetics
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Summary:Hepatitis C virus (HCV) infection is a major worldwide health problem, causing chronic hepatitis, liver cirrhosis and primary liver cancer. In addition to its role in the viral polyprotein-processing, the viral NS3 serine protease has been implicated in interactions with various cell constituents resulting in phenotypic changes including malignant transformation. NS3 is currently regarded a prime target for anti-viral drugs thus specific inhibitors of its activities should be of importance. With the aim of inhibiting NS3-mediated cell transformation, we used antibody-phage display to isolate NS3-specific single-chain antibodies (scFv). We have isolated and characterized eight anti-NS3 scFvs. We investigated the phenotypic changes that NS3-expressing cells undergo upon intracellular expression of these antibodies in different subcellular compartments (intracellular immunization), assayed by their proliferation rate and their ability to grow anchorage independently. The intracellular location of NS3 and the scFvs were analyzed by immunofluorescent staining using confocal microscopy. Nuclear targeted anti-NS3 intrabodies shuttle NS3 from the cytosol to the nucleus with concomitant inhibition of cell proliferation and loss of the transformed phenotype. Intracellular immunization-based gene therapy strategies may emerge as a promising antiviral approach to interfere with the life cycle and tumorigenicity of HCV.
ISSN:0168-8278
1600-0641
DOI:10.1016/j.jhep.2004.02.026