NF-κB plays a key role in inducing CD274 expression in human monocytes after lipopolysaccharide treatment.
NF-κB plays a key role in inducing CD274 expression in human monocytes after lipopolysaccharide treatment.
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DOI：
10.1371/journal.pone.0061602
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Bai Y
中科院分区：
文献类型：
--
作者：
Huang G;Wen Q;Zhao Y;Gao Q;Bai Y
CD274 是程序性死亡 1 的两种共刺激配体之一，在单核吞噬细胞系统 (MPS) 中广泛表达，可能共同刺激 T 细胞并调节炎症反应。然而，人们对炎症反应过程中 CD274 基因表达的变化及其潜在分子机制知之甚少。因此，描述调节 CD274 表达的复杂机制对于理解炎症反应期间的免疫调节系统至关重要。本研究的目的是评估体外单核细胞炎症反应模型中调节 CD274 表达的分子机制。首先，观察脂多糖（LPS）处理后人原代单核细胞中CD274的表达水平并与NF-κB激活相关。其次，根据假定的 NF-κB 结合位点的分布，构建了 5' 截短的人 CD274 启动子报告基因，转染至 U937 细胞中，并通过双荧光素酶测定鉴定了基础（nt -570 至 +94）和 LPS 诱导（nt -1735 至 -570）转录的关键启动子区域。最后，通过点突变分析和染色质免疫沉淀分析（ChIP）对 LPS 诱导的 CD274 转录活性的关键 NF-κB 结合位点（nt -610 至 -601）进行了表征。因此，本研究为理解某些感染和炎症性疾病中 CD274 表达的控制机制奠定了分子基础。
CD274, one of two co-stimulatory ligands for programmed death 1 and widely expressed in the mononuclear phagocyte system (MPS), may co-stimulate T cells and regulates inflammatory responses. However, changes in CD274 gene expression and the underlying molecular mechanism are poorly understood during inflammatory responses. Therefore, delineation of the complex mechanisms regulating CD274 expression is critical to understand this immunoregulatory system during inflammatory responses. The purpose of this study was to assess the molecular mechanisms regulating CD274 expression in an in vitro monocyte model of inflammatory response. Firstly, CD274 expression levels in human primary monocytes after lipopolysaccharide (LPS) treatment were observed and correlated with NF-κB activation. Secondly, based on the distribution of putative NF-κB binding sites, 5′ truncated human CD274 promoter reporters were constructed, transfected into U937 cells and critical promoter regions for basal (nt −570 to +94) and LPS-induced (nt −1735 to −570) transcription were identified by dual luciferase assays. Finally, a key NF-κB binding site (nt −610 to −601) for LPS-inducible CD274 transcriptional activity was characterized by point mutation analysis and chromatin immunoprecipitation analysis assays (ChIP). Thus, the present study establishes a molecular basis to understand the mechanisms governing CD274 expression in certain infections and inflammatory disorders.
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影响因子：
15.9
作者：
Lien, E;Means, TK;Golenbock, DT
通讯作者：
Golenbock, DT
DOI：
10.1016/j.injury.2007.10.003
发表时间：
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