Large-scale production and purification of a vaccinia recombinant-derived HIV-1 gp160 and analysis of its immunogenicity.
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.
Barrett
N
N
Immuno AG, Vienna, Austria.
Mitterer
A
A
Mundt
W
W
Eibl
J
J
Eibl
M
M
Gallo
R C
RC
Moss
B
B
Dorner
F
F
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
D000276:Adjuvants, Immunologic; D019302:HIV Envelope Protein gp160; D012191:Retroviridae Proteins; D014612:Vaccines; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins