title: Induction of hepatitis C virus E1 envelope protein-specific immune response can be enhanced by mutation of N-glycosylation sites.
abstract: Deglycosylation of viral glycoproteins has been shown to influence the number of available epitopes and to modulate immune recognition of antigens. We investigated the role played by N-glycans in the immunogenicity of hepatitis C virus (HCV) E1 envelope glycoprotein, a naturally poor immunogen. Eight plasmids were engineered, encoding E1 protein mutants in which the four N-linked glycosylation sites of the protein were mutated separately or in combination. In vitro expression studies showed an influence of N-linked glycosylation on expression efficiency, instability, and/or secretion of the mutated proteins. Immunogenicity of the E1 mutants was studied in BALB/c mice following intramuscular and intraepidermal injection of the plasmids. Whereas some mutations had no or only minor effects on the antibody titers induced, mutation of the fourth glycosylation site (N4) significantly enhanced the anti-E1 humoral response in terms of both seroconversion rates and antibody titers. Moreover, antibody induced by the N4 mutant was able to recognize HCV-like particles with higher titers than those induced by the wild-type construct. Epitope mapping indicated that the E1 mutant antigens induced antibody directed at two major domains: one, located at amino acids (aa) 313 to 332, which is known to be reactive with sera from HCV patients, and a second one, located in the N-terminal domain of E1 (aa 192 to 226). Analysis of the induced immune cellular response confirmed the induction of gamma interferon-producing cells by all mutants, albeit to different levels. These results show that N-linked glycosylation can limit the antibody response to the HCV E1 protein and reveal a potential vaccine candidate with enhanced immunogenicity.
authors.affiliation: Unité Mixte CNRS/BioMérieux, 69364 Lyon Cédex 07, France.
authors.forename: A
authors.initials: A
authors.lastname: Fournillier
authors.forename: C
authors.initials: C
authors.lastname: Wychowski
authors.forename: D
authors.initials: D
authors.lastname: Boucreux
authors.forename: T F
authors.initials: TF
authors.lastname: Baumert
authors.forename: J C
authors.initials: JC
authors.lastname: Meunier
authors.forename: D
authors.initials: D
authors.lastname: Jacobs
authors.forename: S
authors.initials: S
authors.lastname: Muguet
authors.forename: E
authors.initials: E
authors.lastname: Depla
authors.forename: G
authors.initials: G
authors.lastname: Inchauspé
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D005260:Female; D006031:Glycosylation; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000145:classification; D007371:Interferon-gamma / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009154:Mutation; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000276:immunology*; D014761:Viral Hepatitis Vaccines / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; C087035:E1 protein, Hepatitis C virus; D007074:Immunoglobulin G; D014759:Viral Envelope Proteins; D014761:Viral Hepatitis Vaccines; D007371:Interferon-gamma