title: Anti-human immunodeficiency virus type 1 activity of phosphorothioate analogs of oligodeoxynucleotides: penetration and localization of oligodeoxynucleotides in HIV-1-infected MOLT-4 cells.
abstract: Phosphorothioate antisense oligodeoxynucleotide against HIV-1 rev (S-ODN-rev) inhibits virus-induced cytopathic effects (CPE) in acute infection and inhibits the expression of HIV-1 core protein, p24, in chronically infected cells in vitro. HIV-1 reverse transcriptase activity was not affected by S-ODN-rev at the high concentrations of 5-25 microM, which were 250-1250 times higher than the concentration required to achieve 100% HIV-1-induced CPE inhibition. [32P]-labeled S-ODN-rev was rapidly uptaken by MOLT-4 cells, whereas [32P]-SO-ODN-rev and [32P]-O-ODN-rev were not. In the observation of FITC-S-ODN-rev-treated MOLT-4 cells by a confocal laser scanning microscope, diffuse fluorescence was apparently observed in the cytoplasm. Interestingly, fluorescence signals were accumulated in the nuclear region of chronically infected MOLT-4/HIV-1 cells 60 min after incubation. FITC-labeled homooligomer, FITC-S-dC20 and FIT-C-S-dT20, also accumulated in the nucleus of MOLT-4/HIV-1 cells, but weak fluorescence was observed on the cell membrane and in the cytoplasm of the FITC-S-random treated MOLT-4/HIV-1 and MOLT-4 cells.
authors.affiliation: Department of Microbiology, Yamanashi Medical University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Nakashima
authors.forename: Y
authors.initials: Y
authors.lastname: Shoji
authors.forename: S G
authors.initials: SG
authors.lastname: Kim
authors.forename: J
authors.initials: J
authors.lastname: Shimada
authors.forename: Y
authors.initials: Y
authors.lastname: Mizushima
authors.forename: M
authors.initials: M
authors.lastname: Ito
authors.forename: N
authors.initials: N
authors.lastname: Yamamoto
authors.forename: H
authors.initials: H
authors.lastname: Takaku
mesh_terms: D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000378:metabolism; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D003593:Cytoplasm / Q000378:metabolism; D015757:Genes, rev / Q000235:genetics; D016655:HIV Core Protein p24 / Q000032:analysis; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000494:pharmacology*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D013601:T-Lymphocytes / Q000821:virology; D013873:Thionucleotides / Q000138:chemical synthesis / Q000494:pharmacology*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D016655:HIV Core Protein p24; D016376:Oligonucleotides, Antisense; D013873:Thionucleotides; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase