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).