Human conjunctival epithelial cells express functional Toll-like receptor 5
Human conjunctival epithelial cells express functional Toll-like receptor 5
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DOI：
10.1136/bjo.2007.128322
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发表时间：
2008-03-01
影响因子：
4.1
通讯作者：
Kinoshita, S.
中科院分区：
文献类型：
--
作者：
Kojima, K.;Ueta, M.;Kinoshita, S.
目的：分析toll样受体5 （TLR5）在人结膜上皮细胞（HCjEC）中的表达及功能。方法：采用逆转录酶（RT） PCR和流式细胞术检测TLR5在HCjEC中的表达。ELISA法测定各组大鼠白细胞介素（IL） 6、IL-8蛋白含量。采用定量RT-PCR检测铜绿假单胞菌、粘质沙雷菌、鼠伤寒沙门菌和枯草芽孢杆菌鞭毛蛋白刺激下信使RNA的表达。免疫组织化学检测TLR5蛋白在人结膜上皮中的定位。结果：HCjEC表达TLR5特异性mRNA和TLR5蛋白。在铜绿假单胞菌和黏结假单胞菌鞭毛蛋白刺激的HCjEC中，IL-6和IL-8的产生增加，IL-6和IL-8 mRNA的表达上调。鼠伤寒沙门氏菌和枯草芽孢杆菌的鞭毛蛋白没有引起这些基因和蛋白的上调。TLR5蛋白在人结膜上皮的基底外侧检测到，而在根尖侧未检测到。结论：人结膜上皮含有功能性TLR5。考虑到TLR5蛋白在基底外侧的空间选择性定位，我们假设当上皮屏障的破坏允许鞭毛蛋白迁移到基底外侧时，而不是在眼表健康的生理条件下，来自眼致病菌的鞭毛蛋白会诱导炎症反应。
Purpose: The expression and function of Toll-like receptor 5 (TLR5) was analysed in human conjunctival epithelial cells (HCjEC).Methods: The expression of TLR5 in HCjEC was studied by reverse transcriptase (RT) PCR and flow cytometry. The amount of interleukin (IL) 6 and IL-8 proteins was determined by ELISA. Messenger RNA expression elicited by stimulation with flagellins derived from Pseudomonas aeruginosa, Serratia marcescens, Salmonella typhimurium, and Bacillus subtilis was assayed by quantitative RT-PCR. The localisation of TLR5 protein in human conjunctival epithelium was detected immunohistochemically.Results: HCjEC expressed TLR5-specific mRNA and TLR5 protein. In HCjEC stimulated with flagellins derived from P aeruginosa and S marcescens, IL-6 and IL-8 production was increased and IL-6 and IL-8 mRNA was upregulated. Flagellins from S typhimurium and B subtilis did not induce the upregulation of these genes and proteins. TLR5 protein was detected on the basolateral but not the apical side of human conjunctival epithelium.Conclusions: Human conjunctival epithelium harbours functional TLR5. Considering the spatially selective basolateral localisation of TLR5 protein, it was postulated that flagellins from ocular pathogenic bacteria induce inflammatory responses when disruption of the epithelial barrier permits their transmigration to the basolateral side but not under healthy physiological conditions on the ocular surface.