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

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