       Document 0233
 DOCN  M9440233
 TI    Quantitation by competitive PCR of HIV-1 proviral DNA in epidermal
       Langerhans cells of HIV-infected patients.
 DT    9404
 AU    Cimarelli A; Zambruno G; Marconi A; Girolomoni G; Bertazzoni U;
       Giannetti A; Istituto di Genetica Biochimica ed Evoluzionistica, CNR,
       Pavia,; Italy.
 SO    J Acquir Immune Defic Syndr. 1994 Mar;7(3):230-5. Unique Identifier :
       AIDSLINE MED/94149556
 AB    Langerhans cells (LC) belong to the dendritic cell family and represent
       the principal antigen presenting cells populating squamous epithelia. We
       have reported the presence of human immunodeficiency virus Type 1
       (HIV-1) proviral DNA and RNA in purified LC from the epidermis of
       seropositive patients. The aim of this study was to quantify HIV-1
       proviral DNA in LC of infected patients using a competitive polymerase
       chain reaction (PCR) assay. Bulk epidermal cell (EC) suspensions were
       obtained from the skin of nine AIDS patients and six seronegative
       subjects. Purified LC and LC-depleted EC were prepared by immunomagnetic
       separation using an anti-CD1a monoclonal antibody. LC preparations did
       not contain T cells, as assessed by reverse transcription PCR analysis
       of the T cell receptor beta-chain gene (C region). In addition, no CD14+
       cells could be detected in LC fractions by immunostaining of cytospin
       preparations. To quantify HIV-1 DNA, a new competitive PCR system was
       devised using SK145/150 as primers (gag) and a competitor plasmid DNA
       with a modified sequence (209 instead of 142 bp). The number of HIV-1
       DNA copies found in the LC of AIDS patients ranged from 107 to
       3,645/10(5) LC. In contrast, LC-depleted EC from the same subjects were
       all negative. The results indicate that in AIDS patients the frequency
       of infected LC is comparable to that reported for peripheral blood CD4+
       T cells.
 DE    Acquired Immunodeficiency Syndrome/*MICROBIOLOGY/PATHOLOGY  Adult  Aged
       Binding, Competitive  DNA, Viral/*ANALYSIS  Female  Human
       HIV-1/GENETICS/*ISOLATION & PURIF  HLA-DR Antigens/ANALYSIS
       Immunomagnetic Separation  Langerhans Cells/*MICROBIOLOGY  Male  Middle
       Age  Plasmids  Polymerase Chain Reaction  Proviruses/GENETICS/*ISOLATION
       & PURIF  Receptors, Antigen, T-Cell, alpha-beta/GENETICS  Rosette
       Formation  Support, Non-U.S. Gov't  Transcription, Genetic  JOURNAL
       ARTICLE

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

