title: A point mutation at Asn-534 that disrupts a conserved N-glycosylation motif of the E2 glycoprotein of hepatitis C virus markedly enhances the sensitivity to antibody neutralization.
abstract: The molecular basis of antibody neutralization against hepatitis C virus (HCV) is poorly understood. The E2 glycoprotein of HCV is critically involved in viral infectivity through specific binding to the principal virus receptor component CD81, and is targeted by anti-HCV neutralizing antibodies. A previous study showed that a mutation at position 534 (N534H) within the sixth N-glycosylation motif of E2 of the J6/JFH1 strain of HCV genotype 2a (HCV-2a) was responsible for more efficient access of E2 to CD81 so that the mutant virus could infect the target cells more efficiently. The purpose of this study was to analyze the sensitivity of the parental J6/JFH1, its cell culture-adapted variant P-47 possessing 10 amino acid mutations and recombinant viruses with the adaptive mutations to neutralization by anti-HCV antibodies in sera of HCV-infected patients. The J6/JFH1 virus was neutralized by antibodies in sera of patients infected with HCV-2a and -1b, with mean 50% neutralization titers being 1:670 and 1:200, respectively (P < 0.00001). On the other hand, the P-47 variant showed 50- to 200-times higher sensitivity to antibody neutralization than the parental J6/JFH1 without genotype specificity. The N534H mutation, and another one at position 416 (T416A) near the first N-glycosylation motif to a lesser extent, were shown to be responsible for the enhanced sensitivity to antibody neutralization. The present results suggest that the residues 534, and 416 to a lesser extent, of the E2 glycoprotein are critically involved in the HCV infectivity and antibody neutralization.
authors.affiliation: Division of Microbiology, Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Mikiko
authors.initials: M
authors.lastname: Sasayama
authors.forename: Ikuo
authors.initials: I
authors.lastname: Shoji
authors.forename: Myrna
authors.initials: M
authors.lastname: Adianti
authors.forename: Da-Peng
authors.initials: DP
authors.lastname: Jiang
authors.forename: Lin
authors.initials: L
authors.lastname: Deng
authors.forename: Takafumi
authors.initials: T
authors.lastname: Saito
authors.forename: Hisayoshi
authors.initials: H
authors.lastname: Watanabe
authors.forename: Sumio
authors.initials: S
authors.lastname: Kawata
authors.forename: Chie
authors.initials: C
authors.lastname: Aoki
authors.forename: Hak
authors.initials: H
authors.lastname: Hotta
mesh_terms: D020816:Amino Acid Motifs; D057134:Antibodies, Neutralizing / Q000097:blood / Q000276:immunology; D002460:Cell Line; D006031:Glycosylation; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology* / Q000472:pathogenicity; D018937:Hepatitis C Antibodies / Q000097:blood / Q000276:immunology*; D006801:Humans; D017354:Point Mutation; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D018937:Hepatitis C Antibodies; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus