       Document 0689
 DOCN  M9640689
 TI    Detection of intracellular HIV-1 Rev protein by flow cytometry.
 DT    9604
 AU    Rigg RJ; Dando JS; Escaich S; Plavec I; Bohnlein E; Progenesys, Palo
       Alto, CA 94304, USA.
 SO    J Immunol Methods. 1995 Dec 27;188(2):187-95. Unique Identifier :
       AIDSLINE MED/96144820
 AB    The Rev trans-activator protein plays a pivotal role in human
       immunodeficiency virus type 1 (HIV-1) replication by allowing expression
       of the viral structural proteins. We have developed a protocol to
       quantitatively assay intracellular steady state levels of Rev Ag (Rev
       wild type and RevM10 proteins) by flow cytometry. Three fixation and
       permeabilization techniques were compared. These protocols varied in the
       magnitude of the signal which could be detected, and in the ability to
       distinguish between Rev Ag positive and negative populations. This
       technology is applicable to a variety transduced or transfected cell
       types (species, lineage), and for cell lines and primary cells acutely
       infected with HIV-1. The assay is therefore a valuable tool both to
       analyze Rev protein expression levels in HIV-infected cells and to
       optimize delivery of the dominant-negative RevM10 gene for clinical gene
       therapy applications. In addition, a second, independent intracellular
       protein (HIV-Tat) has been detected using the same approach.
 DE    Animal  Antibodies, Monoclonal  Cell Line  Cells, Cultured  Comparative
       Study  Flow Cytometry/*METHODS  Gene Products, rev/*ANALYSIS  Gene
       Products, tat/ANALYSIS  Hela Cells/VIROLOGY  Human
       HIV-1/*CHEMISTRY/PHYSIOLOGY  Lymphocytes/VIROLOGY  Mice  Permeability
       Reproducibility of Results  Tissue Fixation  Transfection  3T3
       Cells/VIROLOGY  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

