Epidermal Langerhans Cell-Targeted Gene Expression by a Dectin-2 Promoter1
Epidermal Langerhans Cell-Targeted Gene Expression by a Dectin-2 Promoter1
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Dectin-2 启动子的表皮朗格汉斯细胞靶向基因表达1
作者：
M. Bonkobara;Paul K. Zukas;S. Shikano;Shin'ichiro Nakamura;P. Cruz;K. Ariizumi
尽管树突状细胞(DC)和朗格汉斯细胞(LC)在初始免疫应答中具有重要的APC功能，但它们很少被用作基因操作的靶点，因为尚未确定控制LC/DC特异性表达的明确调控元件。此前，我们发现Dectin-2是一种C型凝集素受体，在类LC的XS细胞系和小鼠表皮内的LC中选择性表达。由于这些特性增加了Dectin-2启动子可能指导LC/DC特异性基因表达的可能性，我们从Dectin-2基因5‘侧翼区(Dec2FR)克隆了一个3.2kb的核苷酸片段，并利用荧光素酶(Luc)报告系统分析了其调控元件和转录活性。Dec2FR包含一个假定的TATA盒和顺式作用元件，如干扰素刺激的反应元件，这些元件驱动XS细胞中特异的基因表达。Dec2FR包括抑制子、增强子和启动子区域，后两个区域共同调节XS细胞特异性基因的表达。在携带Dec2FR调节的Luc基因的转基因小鼠中，皮肤是Luc活性的主要部位，LC是表皮内这种活性的唯一来源。相比之下，其他APC(DC、巨噬细胞和B细胞)和T细胞表达的Luc活性接近背景水平。我们的结论是，在体外和体内，Dec2FR选择性地靶向于高水平的构成基因的表达。我们的发现为在LC靶向基因表达系统中使用Dectin-2启动子以增强疫苗效力和调节免疫反应奠定了基础。
Despite their critical function as APCs for primary immune responses, dendritic cells (DC) and Langerhans cells (LC) have been rarely used as targets of gene-based manipulation because well-defined regulatory elements controlling LC/DC-specific expression have not been identified. Previously, we identified dectin-2, a C-type lectin receptor expressed selectively by LC-like XS cell lines and by LC within mouse epidermis. Because these characteristics raised the possibility that dectin-2 promoter may direct LC/DC-specific gene expression, we isolated a 3.2-kb nucleotide fragment from the 5′-flanking region of the dectin-2 gene (Dec2FR) and characterized its regulatory elements and the transcriptional activity using a luciferase (Luc) reporter system. The Dec2FR contains a putative TATA box and cis-acting elements, such as the IFN-stimulated response element, that drive gene expression specifically in XS cells. Dec2FR comprises repressor, enhancer, and promoter regions, and the latter two regions coregulate XS cell-specific gene expression. In transgenic mice bearing a Dec2FR-regulated Luc gene, the skin was the predominant site of Luc activity and LC were the exclusive source of such activity within epidermis. By contrast, other APCs (DC, macrophages, and B cells) and T cells expressed Luc activity close to background levels. We conclude that epidermal LC are targeted selectively for high-level constitutive gene expression by Dec2FR in vitro and in vivo. Our findings lay the foundation for use of the dectin-2 promoter in LC-targeted gene expression systems that may enhance vaccination efficacy and regulate immune responses.
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从鼠表皮连续产生抗原呈递树突状细胞系。
DOI：
--
发表时间：
1995
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Xu,S;Ariizumi,K;Caceres-Dittmar,G;Edelbaum,D;Hashimoto,K;Bergstresser,PR;Takashima,A
通讯作者：
Takashima,A
小鼠口服接种后，主要在肠道相关树突状细胞中检测到水基微胶囊。
影响因子：
5.5
作者：
Lomotan,EA;Brown,KA;Speaker,TJ;Offita,PA
通讯作者：
Offita,PA
体内肠上皮内淋巴细胞和 CD8 T 细胞上抗原驱动的 CD11c 诱导。
DOI：
--
发表时间：
1995
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Huleatt,JW;Lefrancois,L
通讯作者：
Lefrancois,L
角质形成细胞中 IFN-γ 依赖性 IL-7 基因调控。
DOI：
--
发表时间：
1995
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Ariizumi,K;Meng,Y;Bergstresser,PR;Takashima,A
通讯作者：
Takashima,A
影响因子：
4.3
作者：
INABA, K;SWIGGARD, WJ;STEINMAN, RM
通讯作者：
STEINMAN, RM