Inhibition of Human Immunodeficiency Virus Type-1 by Retroviral Vectors Expressing Antisense-TAR
- 1 December 1994
- journal article
- research article
- Published by Mary Ann Liebert Inc in Human Gene Therapy
- Vol. 5 (12) , 1467-1475
- https://doi.org/10.1089/hum.1994.5.12-1467
Abstract
The human immunodeficiency virus type-1 (HIV-1) Tat activation response (TAR) region is essential for Tat-mediated trans-activation of the HIV-1 long terminal repeat (LTR). The TAR element is present on the 5′ and 3′ ends of all HIV-1 transcripts and is relatively conserved among different HIV-1 isolates. These properties make it an attractive target for anti-HIV-1 gene therapy strategies. We have constructed a Moloney murine leukemia-based retroviral vector that expresses a chimeric tRNAiMet-antisense TAR fusion transcript complementary to the HIV-1 TAR region. The potential of this anti-TAR retroviral vector to inhibit HIV-1 was initially tested by transient transfections with an HIV-1–LTR–Tat expression plasmid into HeLa–CAT cells. Anti-TAR inhibited Tat-mediated HIV-1 LTR-driven CAT reporter gene expression in a dose-dependent fashion. The antisense-TAR vector was then used to transduce the human SupT1 T cell line. Cotransfection of these SupT1 cells with a Tat expression plasmid plus an HIV-1 LTR–CAT reporter plasmid resulted in decreased CAT gene expression in comparison to control transduced SupT1 cells. The antisense-TAR engineered SupT1 cell line was then challenged with HIV-1MN. HIV-1 viral production was inhibited in SupT1 cells transduced with the antisense-TAR retroviral vector. Greater inhibition of HIV-1 was observed with antisense-TAR as compared to antisense-Tat expressing retroviral vector. These observations suggest that antisense-TAR retroviral vectors are potentially useful for clinical anti-HIV-1 gene therapy. One of the major goals in anti-HIV gene therapy is to develop an effective, antisense-based anti-HIV strategy that specifically targets essential, conserved, and preferably multiple regions on the HIV-1 mRNA. The HIV-1 TAR region meets these criteria because it is essential for Tat trans-activation, is conserved among primary isolates, and is present on both ends of all HIV-1 mRNA and the viral genome itself. A general advantage of antisense-based strategies, compared with trans-dominant viral proteins, is the lack of immunogenicity. Based on these reasons, we developed an antisense-TAR retroviral vector and demonstrate effective inhibition of HIV-1 in vitro. This antisense retroviral vector therefore represents a potential candidate for clinical anti-HIV gene therapy.Keywords
This publication has 28 references indexed in Scilit:
- Activation of HIV transcription by TatCurrent Opinion in Genetics & Development, 1992
- Dual-target Inhibition of HIV-1 in Vitro by Means of an Adeno-Associated Virus Antisense VectorScience, 1992
- Overexpression of TAR sequences renders cells resistant to human immunodeficiency virus replicationCell, 1990
- Retroviral Vectors Expressing Soluble CD4: A Potential Gene Therapy for AIDSAIDS Research and Human Retroviruses, 1990
- Tat trans-activates the human immunodeficiency virus through a nascent RNA targetCell, 1989
- Temporal fluctuations in HIV quasispecies in vivo are not reflected by sequential HIV isolationsCell, 1989
- A Quantitative Bioassay for HIV-1 Based on Trans-ActivationScience, 1988
- Expression of the art/trs Protein of HIV and Study of Its Role in Viral Envelope SynthesisScience, 1987
- Regulation of mRNA accumulation by a human immunodeficiency virus trans-activator proteinCell, 1987
- Expression and Characterization of the Trans -Activator of HTLV-III/LAV VirusScience, 1986