title: [Vaccination with three HIV-1 cross neutralizing epitopes fused to HBV S antigen could induce robust antibody immune response in mice].
abstract: To enhance immunogenicity of HIV-1 cross neutralizing epitopes , three HIV-1 cross neutralizing epitopes (ELDKWA, NWFDIT, GPGRAFY) were fused to 3' end of HBV S gene by PCR cloning technology, respectively. Three vaccinia virus (Tiantan strain) recombinants expressing separately the three fusion genes were subsequently constructed, named as RVJ1175S-2F5 (ELDKWA), RVJ1175S-4E10 (NWFDIT) and RVJ1175S-447-52D (GPGRAFY), respectively. From the supernatants of CEF cells infected by these vaccinia recombinants, three subunit vaccines (PS-2F5, PS-4E10 and PS-447-52D) were prepared after purification. Biology and immunology characteristics of these fusion antigens in vaccinia recombinants and subunit vaccines were comparatively studied. It was confirmed by PCR and sequencing that the fusion genes were inserted into the TK locus of vaccinia virus (Tiantan strain) correctly. The Fusion proteins were expressed efficiently and secreted into supernatant of the infected cells, which was demonstrated by HBsAg ELISA test. Two typical HBsAg bands of 23kD and 27kD were detected in all the purified samples by SDS-PAGE. These two bands were reacted well to HBsAb and corresponding HIV-1 monoclonal antibodies 2F5, 4E10 and 447-52D. BALB/c mice were immunized with subunit and vaccinia recombinant vaccines by intraperitoneal injection. High levels of HBsAb and anti-HIV-1 cross neutralizing epitope antibody in peripheral blood of immunized mice were tested by ELISA, and all the antibody titers induced by three subunit vaccines were higher than that induced by correlated vaccinia recombinants in mice. This work provides a basis for future study on neutralizing activity of these immunized sera and enhancing immune effect through the combined immunization with different type of vaccines.
authors.affiliation: National Institute for Virual Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
authors.forename: Xue-ren
authors.initials: XR
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.forename: Wen
authors.initials: W
authors.lastname: Wang
authors.forename: Yao
authors.initials: Y
authors.lastname: Deng
authors.forename: Xiang-rong
authors.initials: XR
authors.lastname: Qi
authors.forename: Ying-ying
authors.initials: YY
authors.lastname: Gao
authors.forename: Xin
authors.initials: X
authors.lastname: Meng
authors.forename: Wen-jie
authors.initials: WJ
authors.lastname: Tan
authors.forename: Li
authors.initials: L
authors.lastname: Ruan
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D015153:Blotting, Western; D003429:Cross Reactions; D000939:Epitopes; D005260:Female; D015497:HIV-1 / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000276:immunology*
chemical_list: D016915:AIDS Vaccines; D000939:Epitopes; D006514:Hepatitis B Surface Antigens; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic