title: Identification of an immunodominant B cell epitope on the hepatitis C virus nonstructural region defined by human monoclonal antibodies.
abstract: Several EBV-transformed B cell lines (BCL) were obtained from two patients with chronic hepatitis C virus (HCV) infection that secreted IgG class antibodies to the HCV nonstructural Ag c100-3. Two cloned BCL, derived from the same parental line, generated stable cloned lines that secreted up to 20 mg/liter of specific IgG1(kappa). Supernatants from oligoclonal and cloned BCL were also analyzed by immunoblot and all strongly reacted with recombinant polypeptides derived from the putative NS4 region of HCV, c100-3 and 5-1-1 (a 42-amino acid fragment of c100-3), whereas no reaction with the viral nucleoprotein, the NS3 nonstructural protein or the superoxide dismutase moiety of the c100-3 fusion protein could be documented. The fine specificity of these antibodies was also evaluated using overlapping synthetic peptides (20-mers) covering the 5-1-1 sequence. All oligoclonal and clonal IgG displayed high affinity binding to peptides covering residues 120-137 of Chiron's c100-3 sequence at the aminoterminus of 5-1-1. In addition, a minimal B cell epitope, N-VLYREF-C, was defined by human oligoclonal and monoclonal antibodies corresponding to residues 132-137. Interestingly, predominant recognition of the N-terminus of 5-1-1 was also observed in more than 80% of sera from patients with HCV infection. In conclusion, we have successfully produced human B cell cloned lines that secrete abundant quantities of IgG1(kappa)-specific for a polypeptide encoded by the NS4 region of HCV. Such antibodies recognize an immunodominant epitope, relative to this region, located at the N-terminus of the 5-1-1 fragment.
authors.affiliation: Instituto di Clinica delle Malattie Infettive, I.R.C.C.S. Policlinico San Matteo, University of Pavia, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Cerino
authors.forename: M U
authors.initials: MU
authors.lastname: Mondelli
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D002213:Capsid / Q000276:immunology*; D000939:Epitopes / Q000032:analysis*; D016174:Hepacivirus / Q000276:immunology*; D006526:Hepatitis C / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008969:Molecular Sequence Data; D014758:Viral Core Proteins / Q000276:immunology*; D017361:Viral Nonstructural Proteins
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D007074:Immunoglobulin G; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins