title: ZAP-70 restoration in mice by in vivo thymic electroporation.
abstract: Viral and non-viral vectors have been developed for gene therapy, but their use is associated with unresolved problems of efficacy and safety. Efficient and safe methods of DNA delivery need to be found for medical application. Here we report a new monopolar system of non-viral electro-gene transfer into the thymus in vivo that consists of the local application of electrical pulses after the introduction of the DNA. We assessed the proof of concept of this approach by correcting ZAP-70 deficient severe combined immunodeficiency (SCID) in mice. The thymic electro-gene transfer of the pCMV-ZAP-70-IRES-EGFP vector in these mice resulted in rapid T cell differentiation in the thymus with mature lymphocytes detected by three weeks in secondary lymphoid organs. Moreover, this system resulted in the generation of long-term functional T lymphocytes. Peripheral reconstituted T cells displayed a diversified T cell receptor (TCR) repertoire, and were responsive to alloantigens in vivo. This process applied to the thymus could represent a simplified and effective alternative for gene therapy of T cell immunodeficiencies.
authors.affiliation: INSERM U928, Université de la Méditerranée, Parc Scientifique de Luminy, Marseille, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Irla
authors.forename: Murielle
authors.initials: M
authors.lastname: Saade
authors.forename: Adrien
authors.initials: A
authors.lastname: Kissenpfennig
authors.forename: Lionel Franz
authors.initials: LF
authors.lastname: Poulin
authors.forename: Lee
authors.initials: L
authors.lastname: Leserman
authors.forename: Patrice N
authors.initials: PN
authors.lastname: Marche
authors.forename: Evelyne
authors.initials: E
authors.lastname: Jouvin-Marche
authors.forename: François
authors.initials: F
authors.lastname: Berger
authors.forename: Catherine
authors.initials: C
authors.lastname: Nguyen
mesh_terms: D000758:Anesthesia; D000818:Animals; D002454:Cell Differentiation; D004553:Electric Conductivity; D018274:Electroporation / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016130:Immunophenotyping; D008221:Lymphoid Tissue / Q000166:cytology; D051379:Mice; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000166:cytology; D013950:Thymus Gland / Q000166:cytology*; D013997:Time Factors; D014162:Transfection; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000172:deficiency / Q000378:metabolism*
chemical_list: D011948:Receptors, Antigen, T-Cell; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D051746:ZAP-70 Protein-Tyrosine Kinase; C497262:Zap70 protein, mouse