Department of Pathology, School of Medical Sciences, Faculty of Medicine, University of New South Wales, Sydney, NSW, Australia.
Invest Ophthalmol Vis Sci. 2010 May;51(5):2558-67. doi: 10.1167/iovs.09-3923. Epub 2009 Dec 17.
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.
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.
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.
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.
眼色素上皮细胞被认为在急性前葡萄膜炎（AAU）的发病机制中起作用，其中 LPS 激活 Toll 样受体（TLRs）可能作为触发因素。在这项研究中，确定了虹膜色素上皮（IPE）中 LPS 受体的表达。
使用 RT-PCR、流式细胞术、Western blot 和免疫组织化学来研究原发性人 IPE 中 LPS 受体复合物（TLR4、MD-2 和 CD14）的表达。通过多重珠阵列和 ELISA 测量 LPS 处理的 IPE 分泌的细胞因子。通过添加可溶性 CD14 和抗体中和研究来研究 CD14 在调节 LPS 反应中的作用。通过免疫组织化学和 Western blot 分析检查 CD14 在体内的表达。
IPE 在体外表达 TLR4、MD-2 和 CD14，并在受到 LPS 刺激时分泌一系列促炎细胞因子（IL-6、CXCL8、CXCL10、CCL2、CCL4 和 CCL5）。LPS 处理的 IPE 中 CXCL8 的分泌依赖于 CD14 和 TLR4。免疫组织化学检测到 CD68+细胞中的 CD14 以及正常房水中的 CD14。
IPE 细胞表达功能性 LPS 受体复合物，并且能够通过分泌一系列促炎介质来促进眼部炎症。CD14 被鉴定为调节 IPE 中 LPS 反应的关键分子。
