title: Single-domain intrabodies against hepatitis C virus core inhibit viral propagation and core-induced NFκB activation.
abstract: Hepatitis C virus (HCV) core plays a key role in viral particle formation and is involved in viral pathogenesis. Here, constructs for single-domain intrabodies consisting of variable regions derived from mouse mAbs against HCV core were established. Expressed single-domain intrabodies were shown to bind to HCV core, and inhibit the growth of cell culture-produced HCV derived from JFH-1 (genotype 2a) and a TH (genotype 1b)/JFH-1 chimera. Adenovirus vectors expressing intrabodies were also capable of reducing HCV propagation. Intrabody expression did not affect viral entry or genome replication of single-round infectious trans-complemented HCV particles. However, intrabody expression reduced intracellular and extracellular infectious titres in CD81-defective Huh7-25 cells transfected with the HCV genome, suggesting that these intrabodies impair HCV assembly. Furthermore, intrabody expression suppressed HCV core-induced NFκB promoter activity. These intrabodies may therefore serve as tools for elucidating the role of core in HCV pathogenesis.
authors: Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Ryosuke
authors: R
authors: Suzuki
authors: Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.|Department of Biological Science and Technology, Tokyo University of Science, Chiba, Japan.
authors: Kenji
authors: K
authors: Saito
authors: Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Mami
authors: M
authors: Matsuda
authors: Animal Immune and Cell Biology Research Unit, National Institute of Agrobiological Sciences, Ibaraki, Japan.
authors: Mitsuru
authors: M
authors: Sato
authors: Laboratory of Molecular Genetics, The Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors: Yumi
authors: Y
authors: Kanegae
authors: Department of Infectious Diseases, Hamamatsu University School of Medicine, Shizuoka, Japan.
authors: Guoli
authors: G
authors: Shi
authors: Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Koichi
authors: K
authors: Watashi
authors: Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Hideki
authors: H
authors: Aizaki
authors: Department of Biological Science and Technology, Tokyo University of Science, Chiba, Japan.
authors: Joe
authors: J
authors: Chiba
authors: Laboratory of Molecular Genetics, The Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors: Izumu
authors: I
authors: Saito
authors: Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Takaji
authors: T
authors: Wakita
authors: Department of Infectious Diseases, Hamamatsu University School of Medicine, Shizuoka, Japan.
authors: Tetsuro
authors: T
authors: Suzuki
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology*; D045744:Cell Line, Tumor; D018604:Epitope Mapping; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D005838:Genotype; D057809:HEK293 Cells; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology; D022781:Hepatocytes / Q000276:immunology* / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D051379:Mice; D016328:NF-kappa B / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000737:chemistry / Q000276:immunology; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D061905:Single-Domain Antibodies / Q000096:biosynthesis / Q000276:immunology*; D014162:Transfection; D014758:Viral Core Proteins / Q000235:genetics* / Q000276:immunology; D019065:Virus Assembly / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D016328:NF-kappa B; D011994:Recombinant Proteins; D061905:Single-Domain Antibodies; D014758:Viral Core Proteins