Morita A, Ariizumi K, Ritter R, Jester J V, Kumamoto T, Johnston S A, Takashima A
Department of Dermatology, University of Texas Southwestern Medical Center, Dallas 75390, USA.
Gene Ther. 2001 Nov;8(22):1729-37. doi: 10.1038/sj.gt.3301580.
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.
朗格汉斯细胞（LC）是抗原呈递细胞树突状细胞（DC）家族中的皮肤特异性成员，在皮肤细胞免疫反应的启动中起关键作用。在本研究中，我们开发了一种靶向LC的基因治疗形式，旨在建立用于LC功能基因操作的原位方案。Dectin-2是一种独特的C型凝集素，由包括表皮LC在内的DC选择性表达。从小鼠dectin-2基因分离的一个3.2 kb的5'侧翼片段，称为dectin-2启动子（pDec2），在XS系列的表皮来源的DC系中表现出显著的转录活性，但在任何测试的非DC系中均未表现出活性。当使用基因枪将pDec2驱动的荧光素酶基因（pDec2-Luc）或增强型绿色荧光蛋白基因（pDec2-EGFP）递送至小鼠皮肤时，几乎仅在IA+群体（即LC）的表皮区室中观察到相应基因产物的表达。基因枪处理部位的LC表现出成熟DC的特征，并且它们迁移至引流淋巴结，这表明靶向LC的基因表达可能导致免疫反应的发展。事实上，在基因枪介导的pDec2-EGFP质粒递送后，可检测到针对EGFP的细胞免疫反应。这些结果引入了一个新的概念，即通过基因枪介导的DNA递送和DC特异性启动子的组合，可以在原位对LC功能进行基因操作。
