       Document 0102
 DOCN  M9640102
 TI    Antiviral activity and protection of cells against human
       immunodeficiency virus type-1 using an antisense
       oligodeoxyribonucleotide phosphorothioate complementary to the 5'-LTR
       region of the viral genome.
 DT    9604
 AU    Anazodo MI; Salomon H; Friesen AD; Wainberg MA; Wright JA; Manitoba
       Institute of Cell Biology, University of Manitoba,; Winnipeg, Canada.
 SO    Gene. 1995 Dec 12;166(2):227-32. Unique Identifier : AIDSLINE
       MED/96125194
 AB    A COS-like monkey kidney cell line stably transfected with the plasmids
       pCMVgagpol-rre-r with the gag and pol genes, and pCMV rev with the rev
       gene of HIV-1 derived from the cDNA clone BH10, was used as a model for
       assessing the effectiveness of antisense (AS) constructs, A 20-mer
       oligodeoxyribonucleotide (oligo) phosphorothioate sequence (5'-CCG CCC
       CTC GCC TCT TGC CG) complementary to a portion of the 5'-long terminal
       repeat (5'-LTR) of the HIV-1 genome was tested for its inhibitory
       effects on the biologically important processes of HIV-1 replication and
       proliferation. We observed a concentration-dependent inhibition of HIV
       protein synthesis. Desitometric analysis of data from Western blot
       analysis showed sequence-specific and concentration-dependent oligo
       inhibition of p24 viral core antigen formation in the low-microM range.
       When lipofectin was used as a delivery vehicle, a markedly increased
       potentiation of the AS activity of the sequence was observed at a lower
       concentration (0.1 microM), following a 24-h preincubation. The AS
       construct specifically inhibited intracellular p24 production in
       chronically HIV-1-infected cells of lymphoid origin (H9/IIIB cells) by
       95%, resulting in a 15-fold inhibitory effect relative to a similar
       sequence thiolated at only seven single-base positions. A
       concentration-dependent attenuation in the reverse transcriptase
       activity and a reduction in viral p24 level was observed in the culture
       supernatant of AS-pretreated HIV-1-infected phytohemagglutinin
       A-stimulated human cord blood mononuclear cells. Incubation of a
       HIV-1-infected lymphoid cell line with AS sequence resulted in a marked
       reduction in syncytium formation, and therefore protected cells from the
       cytopathic effects of the virus. Furthermore, the AS oligo did not
       appear to be cytotoxic in cell growth rate and colony-forming ability
       assays. The AS oligo described in this report is a useful new tool for
       the molecular analysis of HIV-1 gene expression and proliferation, and
       may have potential as a therapeutic agent.
 DE    Animal  Base Sequence  Cell Division  Cell Line  Cercopithecus aethiops
       Gene Expression Regulation, Viral  HIV Infections/*PREVENTION & CONTROL
       HIV Long Terminal Repeat/GENETICS  HIV-1/*GENETICS  Molecular Sequence
       Data  Oligonucleotides, Antisense/*THERAPEUTIC USE  RNA-Directed DNA
       Polymerase/GENETICS  Support, Non-U.S. Gov't
       T-Lymphocytes/CYTOLOGY/MICROBIOLOGY  Thionucleotides/THERAPEUTIC USE
       Virus Replication  JOURNAL ARTICLE

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

