title: Usefulness of the phage display technology for the identification of a hepatitis C virus NS4A epitope recognized early in the course of the disease.
abstract: A dodecapeptide phage-displayed library was screened with the mouse monoclonal antibody (mAb) 2E3C2 which competed with human antibodies for the binding to the HCV c100 recombinant protein. Four mimotopes shared a consensus motif with the HCV 1701-1707 sequence corresponding to the carboxyl-terminal domain of the non-structural protein NS4A. However, these mimotopes reacted with 2E3C2 only, whereas the corresponding NS4 epitope defined at the sequence 1698-1709 and displayed on phage was recognized by both 2E3C2 and sera from HCV infected patients. Using the Spot method of multiple peptide synthesis and alanine replacement analysis, the respective reactivities of mAb 2E3C2 and anti-NS4A human antibodies against NS4 were shown to be directed against two slightly different overlapping minimal linear sequences and to involve different critical residues. The phage clone displaying the NS4 epitope was used to study the specific recognition of this epitope by different individual HCV positive sera as well as by two seroconversion panels of sera from HCV infected patients. Compared with the detection by RIBA of the different HCV antigens and c100 particularly, these results indicated that the antibodies directed against the NS4 (1698-1709) epitope were produced early during the course of the disease and decreased later.
authors.affiliation: Unité Mixte de Recherche UMR 2714 CNRS-bioMérieux, IFR 128 BioSciences Lyon Gerland, 21 avenue Tony Garnier, 69365 Lyon Cedex 07, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Ferrieu-Weisbuch
authors.forename: Florence
authors.initials: F
authors.lastname: Bettsworth
authors.forename: Laurence
authors.initials: L
authors.lastname: Becquart
authors.forename: Glaucia
authors.initials: G
authors.lastname: Paranhos-Baccala
authors.forename: Sandrine
authors.initials: S
authors.lastname: Michel
authors.forename: Michel
authors.initials: M
authors.lastname: Arnaud
authors.forename: Colette
authors.initials: C
authors.lastname: Jolivet-Reynaud
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002352:Carrier Proteins / Q000276:immunology*; D018604:Epitope Mapping; D000939:Epitopes / Q000032:analysis*; D005260:Female; D016174:Hepacivirus / Q000276:immunology*; D006526:Hepatitis C / Q000276:immunology*; D018937:Hepatitis C Antibodies / Q000097:blood* / Q000276:immunology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007700:Kinetics; D051379:Mice; D019151:Peptide Library; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D000939:Epitopes; D018937:Hepatitis C Antibodies; D047908:Intracellular Signaling Peptides and Proteins; C455041:NS4A cofactor peptide, Hepatitis C virus; D019151:Peptide Library; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins