Development of a Langerhans cell-targeted gene therapy format using a dendritic cell-specific promoter.
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.
Morita
A
A
Department of Dermatology, University of Texas Southwestern Medical Center, Dallas 75390, USA.
Ariizumi
K
K
Ritter
R
R
Jester
J V
JV
Kumamoto
T
T
Johnston
S A
SA
Takashima
A
A
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
D037102:Lectins; D037181:Lectins, C-Type; D008164:Luminescent Proteins; C445340:dectin-2, mouse; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma; D008156:Luciferases