title: Development of a Langerhans cell-targeted gene therapy format using a dendritic cell-specific promoter.
abstract: Langerhans cells (LC), which are a skin-specific member of the dendritic cell (DC) family of antigen presenting cells, play critical roles in the initiation of cellular immune responses in the skin. We developed a LC-targeted gene therapy format in this study, aimed at the establishment of in situ protocols for genetic manipulation of LC function. Dectin-2 is a unique C-type lectin that is expressed selectively by DC, including epidermal LC. A 3.2 kb 5' flanking fragment isolated from the mouse dectin-2 gene, termed the dectin-2 promoter (pDec2), exhibited significant transcriptional activities in epidermal-derived DC lines of the XS series, but not in any of the tested non-DC lines. When pDec2-driven luciferase gene (pDec2-Luc) or enhanced green fluorescence protein gene (pDec2-EGFP) was delivered to mouse skin using the gene gun, expression of the corresponding gene product was observed in the epidermal compartment almost exclusively by the IA+ population (ie LC). LC in the gene gun-treated sites showed features of mature DC and they migrated to the draining lymph node, suggesting that LC-targeted gene expression may lead to the development of immune responses. In fact, EGFP-specific cellular immune responses became detectable after gene gun-mediated delivery of pDec2-EGFP plasmid. These results introduce a new concept that LC function can be genetically manipulated in situ by the combination of gene gun-mediated DNA delivery and a DC-specific promoter.
authors.affiliation: Department of Dermatology, University of Texas Southwestern Medical Center, Dallas 75390, USA.
authors.forename: A
authors.initials: A
authors.lastname: Morita
authors.forename: K
authors.initials: K
authors.lastname: Ariizumi
authors.forename: R
authors.initials: R
authors.lastname: Ritter
authors.forename: J V
authors.initials: JV
authors.lastname: Jester
authors.forename: T
authors.initials: T
authors.lastname: Kumamoto
authors.forename: S A
authors.initials: SA
authors.lastname: Johnston
authors.forename: A
authors.initials: A
authors.lastname: Takashima
mesh_terms: D000818:Animals; D019470:Biolistics; D002465:Cell Movement; D005260:Female; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma / Q000096:biosynthesis; D007801:Langerhans Cells / Q000276:immunology / Q000378:metabolism*; D037102:Lectins / Q000235:genetics*; D037181:Lectins, C-Type; D008156:Luciferases / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
chemical_list: D037102:Lectins; D037181:Lectins, C-Type; D008164:Luminescent Proteins; C445340:dectin-2, mouse; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma; D008156:Luciferases