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Nucleic Acids Research, 1994, Vol. 22, No. 23 5004-5010
© 1994

MOLECULAR BIOLOGY

Anti-human immunodeficiency virus type 1 activity of phosphorothioate analogs of oligodeoxynucleotides: penetration and localization of oligodeoxynucleotides in HIV-1-infected MOLT-4 cells

Hideki Nakashima*, Yoko Shoji1, Sang-Gun Kim2, Jingoro Shimada1, Yutaka Mizushima1, Masahiko Ito, Naoki Yamamoto3 and Hiroshi Takaku2

Department of Microbiology, Yamanashi Medical University Nakakoma-gun, Yamanashi 409-38 1Institute of Medical Science, St Marianna University School of Medicine Kawasaki, Kanagawa 216 2Department of Industrial Chemistry, Chiba Institute of Technology Tsudanuma, Narashino, Chiba 275 3Department of Microbiology, Tokyo Medical and Dental University School of Medicine Bunkyo-ku, Tokyo 113, Japan

*To whom correspondence should be addressed

Received August 8, 1994. Revised September 30, 1994. Accepted September 30, 1994.

Phosphorothioate antisense oligodeoxynucleotide against HIV-1 rev (S-ODU-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 µM, 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-reiMreated 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.


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W.-S. Park, N. Miyano-Kurosaki, M. Hayafune, E. Nakajima, T. Matsuzaki, F. Shimada, and H. Takaku
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