title: Large-scale production and purification of a vaccinia recombinant-derived HIV-1 gp160 and analysis of its immunogenicity.
abstract: The human immunodeficiency virus (HIV-1) envelope gene was expressed in large-scale microcarrier cultures of Vero cells using a system involving coinfection with two recombinant vaccinia viruses. One recombinant contained the bacteriophage T7 RNA polymerase gene under control of a vaccinia virus promoter. The second contained the HIV-1 gp160 gene flanked by T7 promoter and termination sequences. The protein was expressed on the surface of infected cells, and it was shown to have a molecular weight of 160 kD and to react with gp41 and gp120 specific monoclonal antibodies. After purification by successive affinity and ion-exchange chromatography, the protein was demonstrated to be present in a particulate form with a diameter in the range of 15-30 nm. When injected into goats a high-titer gp160 specific antibody response was elicited and group-specific neutralizing activity could be demonstrated in vitro. The immunogenicity of the protein was also studied in conjunction with a number of adjuvant formulations, and the highest potency in mice was obtained using a preparation with 0.2% Al(OH)3 and 0.25% deoxycholate.
authors.affiliation: Immuno AG, Vienna, Austria.
authors.forename: N
authors.initials: N
authors.lastname: Barrett
authors.forename: A
authors.initials: A
authors.lastname: Mitterer
authors.forename: W
authors.initials: W
authors.lastname: Mundt
authors.forename: J
authors.initials: J
authors.lastname: Eibl
authors.forename: M
authors.initials: M
authors.lastname: Eibl
authors.forename: R C
authors.initials: RC
authors.lastname: Gallo
authors.forename: B
authors.initials: B
authors.lastname: Moss
authors.forename: F
authors.initials: F
authors.lastname: Dorner
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D006041:Goats; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000276:immunology*; D051379:Mice; D009500:Neutralization Tests; D012191:Retroviridae Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D014612:Vaccines / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology* / Q000302:isolation & purification; D014616:Vaccinia virus / Q000235:genetics; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000276:Adjuvants, Immunologic; D019302:HIV Envelope Protein gp160; D012191:Retroviridae Proteins; D014612:Vaccines; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins