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T-cell protection and enrichment through lentiviral CCR5 intrabody gene delivery

Gene Therapy volume 13, pages 1480–1492 (2006)Cite this article

A Corrigendum to this article was published on 20 March 2007

Abstract

CCR5 is the chemokine co-receptor for R5-tropic human immunodeficiency virus type 1 (HIV-1) isolates most often associated with primary infection. We have developed an HIV-1 self-inactivating vector, CAD-R5, containing a CCR5 single-chain antibody (intrabody) gene, which when expressed in T-cell lines and primary CD4+ T cells disrupts CCR5 cell surface expression and provides protection from R5-tropic isolate exposure. Furthermore, CAD-R5 intrabody expression in primary CD4+ T cells supports significant growth and enrichment over time during HIV-1-pulsed dendritic cell–T-cell interactions. These results indicate that CCR5 intrabody-expressing CD4+ T cells are refractory against this highly efficient primary route of infection. CD34+ cells transduced with the CAD-R5 vector gave rise to CD4+ and CD8+ thymocytes in non-obese diabetic (NOD)/ severely combined-immunodeficient (SCID)-human thymus/liver (hu thy/liv) mice, suggesting that CCR5 intrabody expression can be maintained throughout differentiation without obvious cellular effects. CD4+ T cells isolated from NOD/SCID-hu thy/liv mice were resistant to R5-tropic HIV-1 challenge demonstrating the maintenance of protection. Our findings demonstrate delivery of anti-HIV-1 activity through CCR5 intrabodies in primary CD4+ T cells and CD34+ cell-derived T-cell progeny. Thus, gene delivery strategies that provide a selective survival and growth advantage for T effector cells may provide a therapeutic benefit for HIV-1-infected individuals who have failed conventional therapies.

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Acknowledgements

We thank Leslie Romero and Laura Crisa, PhD, MD, TSRI, for assistance with the NOD/SCID-hu thy/liv surgeries and mouse maintenance, Mike McCune, MD, PhD, Cheryl Stoddart, PhD, and Jose Rivera, UCSF, for SCID-hu training, and Jerry Zack, PhD, and Beth Jamieson, PhD, UCLA, for mHSA viruses. We are grateful for the support of the late Dr Nava Sarver. We also thank the UCSD CFAR (5P30 AI36214) for p24 determination. BB was supported by a Fellowship from UARP F00-SRI-036. This research was supported by NIH/NAID AI49165, AI40882 and AI29329-16 (BET) and GM065059 (CFB). This is publication 17294-MEM from The Scripps Research Institute.

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  1. C H Swan and B Bühler: These authors contributed equally to this work.

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  1. Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, USA

    C H Swan, B Bühler, M P Tschan & B E Torbett

  2. Department of Molecular Biology, La Jolla, CA, USA

    C F Barbas III

  3. Department of Molecular Pathology, University of California, San Diego, La Jolla, CA, USA

    C H Swan & B E Torbett

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Swan, C., Bühler, B., Tschan, M. et al. T-cell protection and enrichment through lentiviral CCR5 intrabody gene delivery. Gene Ther 13, 1480–1492 (2006). https://doi.org/10.1038/sj.gt.3302801

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