title: Multimeric HCV E2 protein obtained from Pichia pastoris cells induces a strong immune response in mice.
abstract: Production of immunogenic hepatitis C virus (HCV) envelope proteins will assist in the future development of preventive or therapeutics applications. Only properly folded monomeric E2 protein is able to bind a putative cellular co-receptor CD81, but this interaction may modulate cell immune function. Recombinant E2 proteins, similar to the native form, but lacking undesirable immunoregulatory features, might be promising components of vaccine candidates against HCV. To obtain E2 suitable for structural as well as functional studies, a recombinant E2 variant (E2680) was produced in Pichia pastoris cells. E2680, comprising amino acids 384 to 680 of the HCV polyprotein, was secreted into the culture supernatant in the N-glycosilated form and was mainly composed of disulfide-linked multimers. Both monomeric and oligomeric forms of E2680 were recognized by conformational-sensitive MAb H53. In addition, antibodies in sera from 70% of HCVpositive patients were reactive against E2680. By immunizing E2680 in BALB/c mice, both a specific cellular immune response and anti-E2680 IgG antibody titers of 1:200,000 were induced. Our data suggest that recombinant E2680 could be useful to successfully induce strong anti-HCV immunity.
authors.affiliation: Hepatitis C Department, Biomedical Research, Center for Genetic Engineering and Biotechnology, Ave. 31 e/ 158 y 190, Cubanacán, Playa, Apdo. 6162, Habana 10600, Cuba.
authors.forename: Gillian
authors.initials: G
authors.lastname: Martínez-Donato
authors.forename: Yanelis
authors.initials: Y
authors.lastname: Capdesuñer
authors.forename: Nelson
authors.initials: N
authors.lastname: Acosta-Rivero
authors.forename: Armando
authors.initials: A
authors.lastname: Rodríguez
authors.forename: Juan
authors.initials: J
authors.lastname: Morales-Grillo
authors.forename: Eduardo
authors.initials: E
authors.lastname: Martínez
authors.forename: Marleny
authors.initials: M
authors.lastname: González
authors.forename: Julio C
authors.initials: JC
authors.lastname: Alvarez-Obregon
authors.forename: Santiago
authors.initials: S
authors.lastname: Dueñas-Carrera
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D001483:Base Sequence; D015153:Blotting, Western; D049109:Cell Proliferation; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D006968:Hypersensitivity, Delayed; D051379:Mice; D008807:Mice, Inbred BALB C; D010843:Pichia / Q000235:genetics*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D017931:DNA Primers; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus