Iris Pigment Epithelial Cells Express a Functional Lipopolysaccharide Receptor Complex
Iris Pigment Epithelial Cells Express a Functional Lipopolysaccharide Receptor Complex
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DOI：
10.1167/iovs.09-3923
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发表时间：
2010-05-01
影响因子：
4.4
通讯作者：
Wakefield, Denis
中科院分区：
文献类型：
--
作者：
Chui, Jeanie J. Y.;Li, Monique W. M.;Wakefield, Denis
目的.假设眼色素上皮细胞在急性前葡萄膜炎（AAU）的发病机制中起作用，其中Toll样受体（TLR）的LPS活化可作为触发因素。本研究采用免疫组织化学方法检测了虹膜色素上皮（IPE）细胞内LPS受体的表达。采用RT-PCR、流式细胞术、Western blot和免疫组织化学方法研究LPS受体复合物（TLR 4、MD-2和CD 14）在人IPE中的表达。通过多重珠阵列和ELISA测量LPS处理的IPE的细胞因子分泌。通过添加可溶性CD 14和通过抗体中和研究来研究CD 14在调节LPS应答中的作用。通过免疫组织化学和Western印迹分析检测体内CD 14的表达。IPE在体外表达TLR 4、MD-2和CD 14，并且当用LPS刺激时分泌一组促炎细胞因子（IL-6、CXCL 8、CXCL 10、CCL 2、CCL 4和CCL 5）。LPS处理的IPE的CXCL 8分泌依赖于CD 14和TLR 4。免疫组化法检测虹膜组织中CD 68+细胞中CD 14的表达，Western blot法检测正常房水中CD 14的表达。IPE细胞表达功能性LPS受体复合物，并能够通过分泌一系列促炎介质来促进眼部炎症。CD 14被鉴定为调节IPE中LPS反应的关键分子。（Invest Ophthalmol维斯科学。2010;51：2558-2567）DOI：10.1167/iovs.09-3923
PURPOSE. Ocular pigment epithelial cells are hypothesized to play a role in the pathogenesis of acute anterior uveitis (AAU), where LPS activation of Toll-like receptors (TLRs) may serve as a trigger. In this study, the expression of LPS receptors in iris pigment epithelium (IPE) was determined.METHODS. RT-PCR, flow cytometry, Western blot, and immunohistochemistry were used to investigate the expression of the LPS receptor complex (TLR4, MD-2, and CD14) in primary human IPE. Cytokine secretion by LPS-treated IPE was measured by multiplex bead array and ELISA. The role of CD14 in modulating the LPS response was investigated by addition of soluble CD14 and by antibody neutralization studies. In vivo expression of CD14 was examined by immunohistochemistry and Western blot analysis.RESULTS. IPE expressed TLR4, MD-2, and CD14 in vitro and secreted a panel of proinflammatory cytokines (IL-6, CXCL8, CXCL10, CCL2, CCL4, and CCL5) when stimulated with LPS. CXCL8 secretion by LPS-treated IPE was dependent on CD14 and TLR4. CD14 was detected in CD68+ cells in the iris by immunohistochemistry and in normal aqueous by Western blot analysis.CONCLUSIONS. IPE cells express a functional LPS receptor complex and are capable of promoting ocular inflammation through secretion of an array of proinflammatory mediators. CD14 was identified as a key molecule that modulated the LPS response in IPE. (Invest Ophthalmol Vis Sci. 2010;51:2558-2567) DOI:10.1167/iovs.09-3923