title: Non-viral gene transfer of murine spleen cells achieved by in vivo electroporation.
abstract: Gene electrotranfer is an attractive physical method to deliver genes to target tissues. The aim of this study was to evaluate in vivo gene electrotransfer into spleen, one of the most important lymphoid organ, in order to create a new tool to modulate the immuno-inflammatory system. C57Bl/6 mice were submitted either to intramuscular electrotransfer (IME) as a reference method or to intrasplenic (ISE) gene electrotransfer. In the naked injected plasmids, the CMV promoter controlled the expression of luciferase, secreted alkaline phosphatase, EGFP, or IFNgamma. The ISE optimal electrotransfer conditions were first determined and ISE was found to be an efficient gene transfer method, which can be used to express secreted or intracellular proteins transiently. Although transfected cells were still present in the spleen 30 days after ISE, transfected spleen cells could recirculate since they were detected in extrasplenic locations. Using a T-lymphocyte-specific promoter controlling the expression of EGFP, splenic T cells could be targeted. Finally, it appeared that ISE procedure does not impair by itself the immune response and does not result in a significant production of antibodies directed to the transgenic proteins in C57Bl/6 mice. This strategy constitutes a new method to manipulate the immune response that can be used in various experimental designs.
authors.affiliation: INSERM U430, Hôpital Broussais, Paris, France.
authors.forename: E
authors.initials: E
authors.lastname: Tupin
authors.forename: B
authors.initials: B
authors.lastname: Poirier
authors.forename: M F
authors.initials: MF
authors.lastname: Bureau
authors.forename: J
authors.initials: J
authors.lastname: Khallou-Laschet
authors.forename: R
authors.initials: R
authors.lastname: Vranckx
authors.forename: G
authors.initials: G
authors.lastname: Caligiuri
authors.forename: A-T
authors.initials: AT
authors.lastname: Gaston
authors.forename: J-P
authors.initials: JP
authors.lastname: Duong Van Huyen
authors.forename: D
authors.initials: D
authors.lastname: Scherman
authors.forename: J
authors.initials: J
authors.lastname: Bariéty
authors.forename: J-B
authors.initials: JB
authors.lastname: Michel
authors.forename: A
authors.initials: A
authors.lastname: Nicoletti
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D018274:Electroporation / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism*; D019076:Transgenes
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma; D008156:Luciferases; D000469:Alkaline Phosphatase