title: Production of immunogenic HIV-1 viruslike particles in stably engineered monkey cell lines.
abstract: A proviral fragment from human immunodeficiency virus type 1 (HIV-1) (LAV-1BRU) containing only protein-coding information, was expressed in COS cells using constitutive promoters in transient and stable transfection experiments. The presence of viruslike particles in cell supernatants was verified by Western blot analysis, density gradient centrifugation, and electron microscopy. Transfection of Vero cells with a similar construct employing the human metallothionein promoter led to the isolation of stable cell lines exhibiting inducible viruslike particle expression in response to cadmium chloride treatment. Induction ratios for viruslike particle expression were in excess of 1000-fold with production levels of p24 core antigen as high as 0.6 mg/L per 24 h. HIV-1 viruslike particles were immunogenic in mice, leading to strong envelope and core-specific humoral responses after two immunizations. The development of stable cell lines expressing significant quantities of HIV-1 viruslike particles offers an alternative to the use of live virus vectors for the production and evaluation of particle-based AIDS vaccines.
authors.affiliation: Connaught Centre for Biotechnology Research, Willowdale, Ontario, Canada.
authors.forename: J R
authors.initials: JR
authors.lastname: Haynes
authors.forename: S X
authors.initials: SX
authors.lastname: Cao
authors.forename: B
authors.initials: B
authors.lastname: Rovinski
authors.forename: C
authors.initials: C
authors.lastname: Sia
authors.forename: O
authors.initials: O
authors.lastname: James
authors.forename: G A
authors.initials: GA
authors.lastname: Dekaban
authors.forename: M H
authors.initials: MH
authors.lastname: Klein
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D005260:Female; D015683:Gene Products, gag / Q000096:biosynthesis; D015483:HIV Antibodies / Q000096:biosynthesis / Q000276:immunology; D015488:HIV Antigens / Q000096:biosynthesis*; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000254:growth & development* / Q000276:immunology / Q000648:ultrastructure; D000882:Haplorhini; D006801:Humans; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D014162:Transfection; D014709:Vero Cells; D014758:Viral Core Proteins / Q000096:biosynthesis; D014771:Virion / Q000254:growth & development* / Q000276:immunology / Q000472:pathogenicity / Q000648:ultrastructure; D014776:Virus Cultivation
chemical_list: D015683:Gene Products, gag; D015483:HIV Antibodies; D015488:HIV Antigens; D016655:HIV Core Protein p24; D014758:Viral Core Proteins; D008668:Metallothionein; D012194:RNA-Directed DNA Polymerase