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M9460275.TXT
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1994-06-12
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Document 0275
DOCN M9460275
TI Development and evaluation of an HIV-1 transfection-neutralization
assay.
DT 9408
AU Back NK; Smit L; Hogervorst E; van Wijk AC; Goudsmit J; Tersmette M;
Department of Virology, University of Amsterdam, Academic Medical;
Center, The Netherlands.
SO J Acquir Immune Defic Syndr. 1994 Jun;7(6):531-8. Unique Identifier :
AIDSLINE MED/94231457
AB We developed a transfection-neutralization assay for human
immunodeficiency virus type 1 (HIV-1) infectious molecular clones. In
this assay CD4 negative adherent cells, transfected in microtiter plates
with fixed amounts of proviral DNA of molecular HIV-1 clones, are
cocultivated with CD4 positive T cell lines or primary peripheral blood
mononuclear cells (PBMC) in the presence of anti-HIV-1 sera or
monoclonal antibodies (MAbs). Results obtained with this technique were
reproducible and compared favorably with a conventional cell-free
infection inhibition assay. The transfection-neutralization assay
obviates the need for virus stock preparation and, therefore, is
particularly suitable for the evaluation of HIV-1 clones with slow
replication kinetics and of recombinant chimeric HIV-1 clones inclined
to undergo additional mutations during stock preparation. The potential
value of this assay for the analysis of the specificity of neutralizing
sera and MAbs was demonstrated in experiments with V3 chimeric molecular
clones.
DE Amino Acid Sequence Antibody Specificity Antigenic
Determinants/ANALYSIS Cell Line Cells, Cultured Chimeric
Proteins/CHEMISTRY/GENETICS/IMMUNOLOGY Cloning, Molecular DNA,
Viral/PHYSIOLOGY Evaluation Studies Hela Cells Human HIV
Antibodies/*BLOOD/IMMUNOLOGY HIV Envelope Protein
gp120/CHEMISTRY/GENETICS/IMMUNOLOGY HIV Infections/IMMUNOLOGY
HIV-1/GENETICS/*IMMUNOLOGY Immune Sera/IMMUNOLOGY Leukocytes,
Mononuclear/MICROBIOLOGY Molecular Sequence Data *Neutralization Tests
Peptide Fragments/CHEMISTRY/GENETICS/IMMUNOLOGY
Proviruses/GENETICS/IMMUNOLOGY Reproducibility of Results Sensitivity
and Specificity Sequence Alignment Support, Non-U.S. Gov't
*Transfection Virus Replication/IMMUNOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).