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Prepublished online as a Blood First Edition Paper on January 2, 2003; DOI 10.1182/blood-2002-02-0578.
Submitted February 25, 2002
Accepted December 23, 2002
Specific transgene expression in human and mouse CD4+ cells using lentiviral vectors with regulatory sequences from the CD4 gene
Gilles Marodon, Enguerran Mouly, Emma J Blair, Charlotte Frisen, Francois M Lemoine, and David Klatzmann*
CNRS UMR7087, CERVI, Paris, France
* Corresponding author; email: david.klatzmann{at}chups.jussieu.fr.
Achieving cell-specific expression of a therapeutic transgene via gene transfer vectors represents a major goal for gene therapy. To achieve specific expression of a transgene in CD4+ cells, we have generated lentiviral vectors expressing the eGFP reporter gene under the control of regulatory sequences derived from the CD4 gene: a minimal promoter and the proximal enhancer, with or without the silencer. Both lentiviral vectors could be produced at high titers (>107 i.p/ml) and were used to transduce normal murine hematopoietic stem cells (HSC). Upon reconstitution of RAG-2 deficient mice with transduced HSCs, the specific vectors were efficiently expressed in T-cells, minimally in B-cells and not in immature cells of the bone marrow. Addition of the CD4 gene silencing element in the vector regulatory sequences led to further restriction of eGFP expression into CD4+ T-cells in reconstituted mice, as well as in ex vivo transduced human T-cells. Non-T CD4+ dendritic and macrophage cells derived from human CD34+ cells in vitro, expressed the transgenes of the specific vectors, albeit at lower levels than CD4+ T-cells. Altogether, we have generated lentiviral vectors that allow specific targeting of transgene expression to CD4+ cells after differentiation of transduced mice HSCs and human mature T-cells. Ultimately, these vectors may prove useful for in situ injections for in vivo gene therapy of HIV infection or genetic immunodeficiencies.
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