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A tissue-specific, activation-inducible, lentiviral vector regulated by human CD40L proximal promoter sequences

Gene Therapy volume 18, pages 364–371 (2011)Cite this article

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Abstract

The application of new protocols for gene therapy against monogenic diseases requires the development of safer therapeutic vectors, particularly in the case of diseases in which expression of the mutated gene is subject to fine regulation, as it is with CD40L (CD154). CD40L, the gene mutated in the X-linked hyper-immunoglobulin M syndrome (HIGM1), is tightly regulated to allow surface expression of its product only on T cells stimulated by antigen encounter. Previous studies in an HIGM1 animal model showed that transduction of progenitor cells corrected the syndrome but caused a thymic lymphoproliferative disease because of the unregulated expression of the transgene by constitutive vectors. To develop a tissue-specific, activation-inducible, lentiviral vector (LV) for gene therapy to counter HIGM1, we have constructed two self-inactivating LVs, pCD40L-eGFP and pCD40L-CD40L, regulated by a 1.3 kb fragment of the human CD40L proximal promoter. The expression of pCD40L-eGFP LV is restricted to cells in which mRNA transcripts of the endogenous CD40L gene can be detected. Moreover, the expression of the reporter gene in primary T lymphocytes depends on the activation state of the cells. Remarkably, primary HIGM1 lymphocytes transduced with pCD40L-CD40L LV expressed CD40L only after T-cell stimulation. Therefore, the CD40L-promoter-driven vectors are able to achieve a near-physiological expression pattern that follows very closely that of the endogenous CD40L gene.

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Acknowledgements

We are grateful to Dr Maria Cruz Garcia (La Paz University Hospital, Madrid, Spain), Dr Jorge Gómez-Sirvent (N.S. Candelaria University Hospital, Tenerife, Spain) and to Dr Graham Davies (Great Ormond Street Hospital London, UK) for their invaluable help in providing blood samples from their patients. We thank our colleague, Dr J Trout, for revising the English text. We acknowledge the continuous and generous supply of rIL-2 provided by the National Institutes of Health AIDS Research and Reference Reagent Program (Rockville, MD, USA). This work was supported by grants SAF2009/09818 (to IJM) and PS09/00340 (to FM) from the Spanish Ministry for Science and Innovation and P06-CTS-02112 from the Department of Innovation and Science, Regional Government of Andalusia (to IJM).

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  1. F Martín and I J Molina: These authors share senior authorship.

Authors and Affiliations

  1. Institute of Biopathology and Regenerative Medicine, Center for Biomedical Research, University of Granada, Health-Science Technology Park, Armilla, Granada, Spain

    Z Romero, S Torres, J D Unciti & I J Molina

  2. University of Granada and Andalucian Department of Health, Andalucian Stem Cell Bank, Center for Biomedical Research, Health-Science Technology Park, Armilla, Granada, Spain

    M Cobo, P Muñoz & F Martín

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  1. Z Romero
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  2. S Torres
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  3. M Cobo
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  7. I J Molina
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Correspondence to F Martín.

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Romero, Z., Torres, S., Cobo, M. et al. A tissue-specific, activation-inducible, lentiviral vector regulated by human CD40L proximal promoter sequences. Gene Ther 18, 364–371 (2011). https://doi.org/10.1038/gt.2010.144

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