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Nucleic Acids Research, 1994, Vol. 22, No. 20 4307-4314
© 1994

STRUCTURAL BIOLOGY

Antisense oligonucleotides in solution or encapsulated in immunoliposomes inhibit replication of HIV-1 by several different mechanisms

Olivier Zelphati, Jean-Louis Imbach1, Nathalie Signoret, Gerald Zon2, Bernard Rayner1 and Lee Leserman*

Centre d'lmmunologie, Institut National de la Santé et de la Recherche Médicale-Centre National de la Recherche Scientifique de Marseille-Luminy Case 906 13288 Marseille Cedex 9 1Laboratoire de Chimie Bioorganique, URA CNRS 488, Universite de Montpellier II Place Eugene Bataillon, 34095 Montpellier, Cedex 5, France 2Lynx Therapeutics, Inc. 3832 Bay Center Place, Hayward, CA 94545, USA

*To whom correspondence should be addressed at: CIML, Case 906, 13288 Marseille Cedex 9, France

Received May 23, 1994. Revised August 24, 1994. Accepted August 24, 1994.

Phosphodiester and phosphorothioate oligonucleotides in {alpha} and ß configurations directed against the initiation codon region of the HIV-1 rev gene were evaluated for their ability to inhibit HIV-1 replication in acutely and chronically infected human CEM cells. Encapsulation in antibody-targeted liposomes (immunoliposomes) permitted intracellular delivery and distinction between oligonucleotide-mediated inhibition of viral entry and intracellular effects on viral RNA. Our results are consistent with four mechanisms of antiviral activity for these antisense oligonucleotides: (i) interference with virus-mediated cell fusion by free but not liposome-encapsulated phosphorothioate oligonucleotides of any sequence; (ii) interference with reverse transcription in a sequence non-specific manner by phosphorothioate oligonucleotides in {alpha} and ß configurations; (iii) interference with viral reverse transcription in a sequence-specific and RNase-H-independent manner by {alpha} and ß phosphodiester oligonucleotides; (iv) interference with viral mRNA in a sequence-specific and RNase-H-dependent manner by ß-phosphorothioate oligonucleotides.


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