title: Human combinatorial libraries yield rare antibodies that broadly neutralize hepatitis C virus.
abstract: One way to dissect the antibody response to an invading microorganism is to clone the antibody repertoire from immune donors and subsequently characterize the specific antibodies. Recently, methodological advances have allowed investigations of neutralizing antibodies against hepatitis C virus (HCV) in vitro. We have investigated three human mAbs, previously isolated from an individual infected with HCV of genotype 2b, that are known to cross-react in a binding assay to the envelope E2 protein of genotypes 1a and 1b. We now report that two of them have a neutralizing activity with a breadth not previously observed. Indeed, mAbs 1:7 and A8 recognized E2 from all of the six major genotypes, and they neutralized retroviral pseudoparticles [HCV pseudoparticles (HCVpp)] carrying genetically equally diverse HCV envelope glycoproteins. Importantly, these antibodies were also able to neutralize the cell culture infectious HCV clone JFH-1 in vitro, with IC(50) values of 60 ng/ml and 560 ng/ml, respectively. The conformational epitopes of these two broadly reactive antibodies were overlapping yet distinct and involved amino acid residues in the 523-535 region of E2, known to be important for the E2-CD81 interaction. The third antibody clone, representing a dominant population in the initial screen for these antibodies, was less broadly reactive and was unable to neutralize the genotype 2a infectious clone JFH-1. Our results confirm at the clonal level that broadly neutralizing human anti-HCV antibodies can be elicited and that the region amino acids 523-535 of the HCV envelope glycoprotein E2 carries neutralizing epitopes conserved across all genotypes.
authors.affiliation: Department of Medicine, Center for Molecular Medicine, Karolinska University Hospital Solna, Karolinska Institutet, 171 76 Stockholm, Sweden.
authors.forename: Daniel X
authors.initials: DX
authors.lastname: Johansson
authors.forename: Cécile
authors.initials: C
authors.lastname: Voisset
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Tarr
authors.forename: Mie
authors.initials: M
authors.lastname: Aung
authors.forename: Jonathan K
authors.initials: JK
authors.lastname: Ball
authors.forename: Jean
authors.initials: J
authors.lastname: Dubuisson
authors.forename: Mats A A
authors.initials: MA
authors.lastname: Persson
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000302:isolation & purification; D002460:Cell Line; D020650:Combinatorial Chemistry Techniques; D017124:Conserved Sequence; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics; D015723:Gene Library; D005838:Genotype; D016174:Hepacivirus / Q000145:classification / Q000235:genetics / Q000276:immunology*; D006526:Hepatitis C / Q000235:genetics / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000235:genetics* / Q000302:isolation & purification; D018936:Hepatitis C Antigens / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D015202:Protein Engineering; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus