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HIV-1 proprotein processing as a target for gene therapy

Gene Therapy volume 10, pages 467–477 (2003)Cite this article

Abstract

The central role of endoconvertases and HIV-1 protease (HIV-1 PR) in the processing of HIV proproteins makes the design of specific inhibitors important in anti-HIV gene therapy. Accordingly, we tested native α1 antitrypsin (α1AT) delivered by a recombinant simian virus-40-based vector, SV(AT), as an inhibitor of HIV-1 proprotein maturation. Cell lines and primary human lymphocytes were transduced with SV(AT) without selection and detectable toxicity. Expression of α1AT was confirmed by Northern blotting, immunoprecipitation and immunostaining. SV(AT)-transduced cells showed no evidence of HIV-1-related cytopathic effects when challenged with high doses of HIV-1NL4-3. As measured by HIV-1 p24 assay, SV(AT)-transduced cells were protected from HIV-1NL4-3 at challenge dose of 40 000 TCID50 (MOI = 0.04). In addition, peripheral blood lymphocytes treated with SV(AT) were protected from HIV doses challenge up to 40 000 TCID50 (MOI = 0.04). By Western blot analyses, the delivered α1AT inhibited cellular processing of gp160 to gp120 and decreased HIV-1 virion gp120. SV(AT) inhibited processing of p55Gag as well. Furthermore, high levels of uncleaved p55Gag protein were detected in HIV virus particles recovered from SV(AT)-transduced cells lines and primary lymphocytes. Thus, delivering α1AT using SV(AT) to human lymphocytes strongly inhibits replication of HIV-1, most likely by inhibiting the activities both of the cellular serine proteases involved in processing gp160 and of the aspartyl protease, HIV-1 PR, which cleaves p55Gag. α1AT delivered by SV(AT) may represent a novel and effective strategy for gene therapy to interfere with HIV replication, by blocking a stage in the virus replicative cycle that has until now been inaccessible to gene therapeutic intervention.

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Acknowledgements

The authors appreciate the technical support of Mr Charles Ko and Ms Maria Lamothe. This work was supported by NIH grants A148244 and RR13156.

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  1. Department of Pathology, Jefferson Medical College, Philadelphia, PA, USA

    P Cordelier & D S Strayer

  2. Department of Medicine, University of California, Davis, CA, USA

    M A Zern

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Cordelier, P., Zern, M. & Strayer, D. HIV-1 proprotein processing as a target for gene therapy. Gene Ther 10, 467–477 (2003). https://doi.org/10.1038/sj.gt.3301891

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