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Inhibition of HIV-1 infection by down-regulation of the CXCR4 co-receptor using an intracellular single chain variable fragment against CXCR4

Gene Therapy volume 8, pages 408–418 (2001)Cite this article

Abstract

CXCR4 is the major co-receptor used by X4 strains of human immunodeficiency virus type I (HIV-1). In HIV-1-infected patients, the appearance of X4 strains (T cell line-tropic) correlates with disease progression. Since its discovery, the CXCR4 co-receptor has been a major target for different agents which block its function, such as stromal-derived factor 1α (SDF-1α) and the anti-CXCR4 monoclonal antibody, 12G5. In the present studies, the 12G5 hybridoma was used to construct a single-chain variable antibody fragment (SFv). Murine leukemia virus (MLV) and simian virus 40 (SV40) were utilized as delivery vehicles for the anti-CXCR4 SFv. Intracellular expression of the anti-CXCR4 SFv led to down-regulation of this critical co-receptor, as demonstrated by immunostaining. This effect significantly and specifically protected transduced cells from challenge with HIV-1, as measured by HIV-1 p24 antigen expression. Inhibition of HIV-1 replication was specific for X4 HIV-1 strains as demonstrated by MAGI assays. HeLa-CD4/βgal-CCR5 cells expressing the anti-CXCR4 SFv showed significant inhibition of infectivity by the X4 HIV-1 strain NL4–3, but not with the R5 HIV-1 strain Bal. Thus, this anti-HIV-1 molecular therapy has the potential to inhibit HIV-1 replication and virion spread. Targeting CXCR4 by intracellular immunization could be of additional benefit to certain HIV-1-infected patients on highly active antiretroviral therapy (HAART).

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Acknowledgements

We wish to thank Ms Rita M Victor and Ms Brenda O Gordon for excellent secretarial assistance, Dr Tim Manser for his critical evaluations of the CXCR4 SFv sequence, Dr Joseph Kulkosky for helpful discussions, and Dr Janet S Butel who kindly supplied the pBSV-1, from which all SV40-derived constructs were created. These studies were supported in part by USPHS grants AI41399 and RR13156 to DSS, and AI38666, NS27405 and AI36557 to RJP.

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Authors and Affiliations

  1. Division of Infectious Diseases, Department of Medicine, The Dorrance H Hamilton Laboratories, Center for Human Virology, Philadelphia, PA, USA

    M BouHamdan, M Mukhtar, L-X Duan & RJ Pomerantz

  2. Department of Pathology, Anatomy and Cell Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA, USA

    DS Strayer & D Wei

  3. Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    J Hoxie

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  1. M BouHamdan
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BouHamdan, M., Strayer, D., Wei, D. et al. Inhibition of HIV-1 infection by down-regulation of the CXCR4 co-receptor using an intracellular single chain variable fragment against CXCR4. Gene Ther 8, 408–418 (2001). https://doi.org/10.1038/sj.gt.3301411

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Keywords

  • HIV-1
  • SFv
  • CXCR4
  • gene therapy

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