Functional Analysis of the Thymic Stromal Lymphopoietin Variants in Human Bronchial Epithelial Cells
Functional Analysis of the Thymic Stromal Lymphopoietin Variants in Human Bronchial Epithelial Cells
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DOI：
10.1165/rcmb.2008-0041oc
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发表时间：
2009-03-01
影响因子：
6.4
通讯作者：
Tamari, Mayumi
中科院分区：
文献类型：
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作者：
Harada, Michishige;Hirota, Tomomitsu;Tamari, Mayumi
胸腺基质淋巴细胞生成素（Thymic stromal lymphopoietin，TSLP）是一种IL-7样细胞因子，其触发树突状细胞介导的辅助性T细胞（Th）2炎症反应，并与人类过敏性疾病的发病机制有关。已经报道了两种TSLP剪接变体。为了发现可能导致疾病的功能性遗传变异，我们对人支气管上皮细胞TSLP基因的单核苷酸多态性（SNP）进行了分析。我们通过对36名受试者的基因组DNA测序，调查了TSLP基因上的SNP，并描述了该基因的连锁不平衡。我们使用实时PCR、报告基因分析和酶联免疫吸附测定来检测SNIPS是否对mRNA表达或蛋白质产生功能性影响。我们在TSLP基因中共发现了23个多态性。在正常人支气管上皮细胞（NHBE）中，poly（I：C）（双链RNA）刺激高度诱导与过敏性炎症相关的TSLP的长型（P = 0.0060）。发现长型TSLP启动子区域的SNIP rs3806933（-847 C> T）产生转录因子激活蛋白（AP）-1的结合位点，并且体外功能分析证明SNP增强AP-1与调节元件的结合。功能性变体增加了长型TSLP对NHBE中poly（I：C）刺激的启动子-报告基因活性。TSLP基因的功能性遗传多态性似乎有助于通过支气管上皮细胞对病毒性呼吸道感染的应答产生更高的TSLP来促进Th 2极化免疫。
Thymic stromal lymphopoietin (TSLP) is an IL-7-like cytokine that triggers dendritic cell-mediated T helper (Th)2 inflammatory responses, and is implicated in the pathogenesis of allergic diseases in humans. Two TSLP splice variants have been reported. To find functional genetic variants that might contribute to disease, we conducted analyses of single nucleotide polymorphisms (SNPs) of the TSLP gene in human bronchial epithelial cells. We surveyed SNPs on the TSLP gene by sequencing genomic DNA from 36 subjects, and characterized the linkage disequilibrium of the gene. We examined whether the SNIPS have functional effects on mRNA expression or protein production using real-time PCR, reporter gene analysis, and enzyme-linked immunosorbent assay. We identified a total of 23 polymorphisms in the TSLP gene. The long form of TSLP, which is associated with allergic inflammation, was highly induced by poly(I:C) (double-stranded RNA) stimulation in normal human bronchial epithelial cells (NHBE) (P = 0.0060). The SNIP rs3806933 (-847C > T) in the promoter region of long-form TSLP was found to create a binding site for the transcription factor activating protein (AP)-1, and in vitro functional analyses demonstrated that the SNP enhanced AP-1 binding to the regulatory element. The functional variant increased promoter-reporter activity of long-form TSLP in response to poly(I:C) stimulation in NHBE. Functional genetic polymorphism of the TSLP gene appears to contribute to Th2-polarized immunity through higher TSLP production by bronchial epithelial cells in response to viral respiratory infections.