Expression of HIV-1 core/envelope chimeric virus-like particles in baculovirus-infected cells.

Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.

Expression of HIV-1 core/envelope chimeric virus-like particles in baculovirus-infected cells.

Abstr Gen Meet Am Soc Microbiol. 1994;94:483 (abstract no. T-7). Unique Identifier : AIDSLINE ASM94/94313085
Truong C; Brand D; Roingeard P; Mallet F; Goudeau A; Barin F; Lab. Virologie, URA CNRS 1334 CHU Bretonneau Tours, France.

Abstract: The viral envelope, involved in the first steps of HIV-1 infection, contains two neutralizing epitopes. The V3 loop induces an early, type-specific HIV-1 neutralizing activity but is highly variable among isolates. Later in infection, broad spectrum neutralizing activity is induced by the conformational CD4 binding site of gp120 (CD4BD env). Moreover, these neutralizing activities are synergic. The core precursor p55 is able of auto assembling into virus-like particles which can be extracellularly released when using the Baculovirus-Insect cells expression system. To obtain recombinant proteins expressing neutralizing epitopes of HIV-1 gp120, several chimeric genes gag-V3env (substitution of different p24 epitopes by a V3 consensus peptide specific of one HIV-1 subtype), gag-CD4BDenv and gag-V3env-CD4BDenv have been constructed by an original procedure using the Polymerase Chain Reaction, and then cloned in a baculovirus transfer vector. After co-transfection of the recombinant vector with baculoviral DNA in SF9 insect cells, recombinant viruses have been selected and purified. Protein expression has been analysed for antigenicity by Western-Blot analysis using several monoclonal antibodies (anti-p17, anti-p24, anti-V3) and by immunoelectron microscopy. All these forms are shown to be antigenic. Formation, budding and extracellular release of virus-like particles have been observed for each construction by electron microscopy on cells dishes as well as on extracellular medium sucrose gradient fractions. Results on immunogenicity should be available to be presented.

Keywords: Animal Antibodies, Monoclonal Baculoviridae Cell Line Chimeric Proteins/ANALYSIS/*BIOSYNTHESIS Cloning, Molecular/METHODS Epitopes Genes, env Genes, gag Genetic Vectors HIV Core Protein p24/ANALYSIS/*BIOSYNTHESIS HIV Envelope Protein gp120/ANALYSIS/*BIOSYNTHESIS HIV-1/GENETICS/*METABOLISM/ULTRASTRUCTURE Moths Polymerase Chain Reaction/METHODS Transfection ABSTRACT
941030
M94A0869

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