Gene-expression analysis of polyI:C-stimulated primary human conjunctival epithelial cells
Gene-expression analysis of polyI:C-stimulated primary human conjunctival epithelial cells
复制标题
DOI：
10.1136/bjo.2010.180554
复制
发表时间：
2010-11-01
影响因子：
4.1
通讯作者：
Kinoshita, Shigeru
中科院分区：
文献类型：
--
作者：
Ueta, Mayumi;Mizushima, Katsura;Kinoshita, Shigeru
研究背景作者先前报道人眼表上皮细胞表达TLR 3，其配体polyI：C刺激IL-6、IL-8和IFN-β的分泌。在这项研究中，研究的综合影响polyI：C刺激的原代人结膜上皮细胞（PHCjECs），作者进行了基因表达分析polyI：C刺激的PHCjECs使用寡核苷酸微阵列，GeneChip.Methods转录上调polyI：C刺激PHCjECs从两个人使用GeneChip检查。11个新的上调的目标转录本，通过定量实时PCR（RT-PCR），并通过ELISA或免疫印迹分析，分别从其他三个individuals.Results基因芯片分析表明，150转录本上调超过3倍，47转录本上调超过10倍后，在PHCjECs的polyI：C刺激，由这些转录本产生的蛋白进行了检查。47个上调转录本中有11个（CXCL11、RIG-I、IL28A、CXCL10、CCL5、CCL4、MDA 5、IL7R、TSLP、CCL20和ICAM-1）在polyI：通过定量RT-PCR检测CXCL 11、CXCL 10、CCL 5、CCL 20、TSLP、结论结膜上皮TLR 3不仅能诱导抗病毒天然免疫应答，而且能调节变态反应。
Background The authors previously reported that human ocular surface epithelium expressed TLR3 and that its ligand polyI:C stimulated the secretion of IL-6, IL-8 and IFN-beta. In this study, to examine comprehensive effects of polyI: C stimulation of primary human conjunctival epithelial cells (PHCjECs), the authors performed a gene-expression analysis of the polyI: C-stimulated PHCjECs using oligonucleotide microarrays, GeneChip.Methods The transcripts upregulated upon polyI: C stimulation in PHCjECs from two individuals were examined using GeneChip. Eleven new upregulated transcripts of interest were confirmed by quantitative real-time PCR (RT-PCR), and seven proteins produced by those transcripts were examined by ELISA or immunoblot analysis in PHCjECs from three other individuals, respectively.Results GeneChip analysis showed that 150 transcripts were upregulated more than threefold and that 47 transcripts were upregulated more than 10-fold upon polyI: C stimulation in the PHCjECs. Eleven of the 47 upregulated transcripts (CXCL11, RIG-I, IL28A, CXCL10, CCL5, CCL4, MDA5, IL7R, TSLP, CCL20 and ICAM-1) were significantly upregulated upon polyI: C stimulation by quantitative RT-PCR, and the levels of seven proteins of the transcripts CXCL11, CXCL10, CCL5, CCL20, TSLP, RIG-I and MDA5 were confirmed by ELISA or immunoblot analysis to increase significantly in polyI: C-stimulated PHCjECs.Conclusions Our results might show that TLR3 of conjunctival epithelium could not only induce antiviral innate immune responses but also regulate the allergic reactions.