Role of N-linked glycans in the functions of hepatitis C virus envelope proteins incorporated into infectious virions.
Hepatitis C virus (HCV) envelope glycoproteins are highly glycosylated, with generally 4 and 11 N-linked glycans on E1 and E2, respectively. Studies using mutated recombinant HCV envelope glycoproteins incorporated into retroviral pseudoparticles (HCVpp) suggest that some glycans play a role in protein folding, virus entry, and protection against neutralization. The development of a cell culture system producing infectious particles (HCVcc) in hepatoma cells provides an opportunity to characterize the role of these glycans in the context of authentic infectious virions. Here, we used HCVcc in which point mutations were engineered at N-linked glycosylation sites to determine the role of these glycans in the functions of HCV envelope proteins. The mutants were characterized for their effects on virus replication and envelope protein expression as well as on viral particle secretion, infectivity, and sensitivity to neutralizing antibodies. Our results indicate that several glycans play an important role in HCVcc assembly and/or infectivity. Furthermore, our data demonstrate that at least five glycans on E2 (denoted E2N1, E2N2, E2N4, E2N6, and E2N11) strongly reduce the sensitivity of HCVcc to antibody neutralization, with four of them surrounding the CD81 binding site. Altogether, these data indicate that the glycans associated with HCV envelope glycoproteins play roles at different steps of the viral life cycle. They also highlight differences in the effects of glycosylation mutations between the HCVpp and HCVcc systems. Furthermore, these carbohydrates form a "glycan shield" at the surface of the virion, which contributes to the evasion of HCV from the humoral immune response.
Helle
François
F
Institut Pasteur de Lille, Bâtiment IBL, 59021 Lille cedex, France.
Vieyres
Gabrielle
G
Elkrief
Laure
L
Popescu
Costin-Ioan
CI
Wychowski
Czeslaw
C
Descamps
Véronique
V
Castelain
Sandrine
S
Roingeard
Philippe
P
Duverlie
Gilles
G
Dubuisson
Jean
J
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D006031:Glycosylation; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006526:Hepatitis C / Q000276:immunology / Q000821:virology*; D006801:Humans; D011134:Polysaccharides / Q000276:immunology / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014771:Virion / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D011134:Polysaccharides; D014759:Viral Envelope Proteins