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M9480860.TXT
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1994-09-05
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Document 0860
DOCN M9480860
TI Expression of HIV-1 core/envelope chimeric virus-like particles in
baculovirus-infected cells.
DT 9410
AU Truong C; Brand D; Roingeard P; Mallet F; Goudeau A; Barin F; Lab.
Virologie, URA CNRS 1334 CHU Bretonneau Tours, France.
SO Abstr Gen Meet Am Soc Microbiol. 1994;94:483 (abstract no. T-7). Unique
Identifier : AIDSLINE ASM94/94313085
AB 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.
DE Animal Antibodies, Monoclonal Antigenic Determinants Baculoviridae
Cell Line Chimeric Proteins/ANALYSIS/*BIOSYNTHESIS Cloning,
Molecular/METHODS 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 MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).