Activation of airway epithelial cells by toll-like receptor agonists
Activation of airway epithelial cells by toll-like receptor agonists
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DOI：
10.1165/rcmb.2003-0388oc
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发表时间：
2004-09-01
影响因子：
6.4
通讯作者：
Schleimer, RP
中科院分区：
文献类型：
--
作者：
Sha, Q;Truong-Tran, AQ;Schleimer, RP
Toll 样受体 (TLR) 在病原体识别和先天免疫中发挥重要作用。我们研究了 BEAS-2B 气道上皮细胞系和原代支气管上皮细胞中 TLR 的存在和功能。标准实时逆转录聚合酶链式反应 (RT-PCR) 分析和 Taqman RT-PCR 显示 BEAS-2B 细胞表达 TLR1-10 的 mRNA。使用有限的微阵列分析测试了几种 TLR 配体激活 BEAS-213 细胞中基因表达的能力，重点关注趋化因子和趋化因子受体家族的基因、细胞因子和信号传导途径。虽然 TLR3 配体双链 RNA 是最有效的上皮激活剂，但也观察到对鞭毛蛋白、脂多糖、CpG、肽聚糖和酵母聚糖的明显反应。 RT-PCR和/或酶联免疫吸附测定用于确认微阵列获得的五种诱导基因的结果：白细胞介素-8、血清淀粉样蛋白A、TLR3、巨噬细胞炎症蛋白-3α和粒细胞-巨噬细胞集落刺激因子。用双链RNA刺激上皮细胞诱导白细胞介素8的水平超过20ng/ml，血清淀粉样蛋白A的水平超过80ng/ml。双链RNA、脂多糖、酵母聚糖A和鞭毛蛋白还诱导巨噬细胞炎症蛋白-3α和粒细胞巨噬细胞集落刺激因子的表达，这可能促进未成熟树突状细胞的迁移和成熟。这些结果表明气道上皮细胞表达多种 TLR，并且它们具有功能活性。 TLR 的上皮表达可能在气道响应吸入病原体的炎症和免疫中发挥重要作用。
Toll-like receptors (TLR) play an important role in pathogen recognition and innate immunity. We investigated the presence and function of TLRs in the BEAS-2B airway epithelial cell line and primary bronchial epithelial cells. Standard real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and Taqman RT-PCR revealed that BEAS-2B cells express mRNA for TLR1-10. Several TLR ligands were tested for their ability to activate gene expression in BEAS-213 cells using limited microarray analyses focusing on genes of the chemokine and chemokine receptor family, cytokines, and signaling pathways. While the TLR3 ligand double-stranded RNA was the most effective epithelial activator, clear responses to flagellin, lipopolysaccharide, CpG, peptidoglycan, and zymosan were also observed. RT-PCR and/or enzyme-linked immunosorbent assay were used to confirm results obtained with microarrays for five of the induced genes: interieukin-8, serum amyloid A, TLR3, macrophage inflammatory protein-3alpha, and granulocyte-macrophage colony-stimulating factor. Stimulation of epithelial cells with double-stranded RNA induced levels of interleukin-8 exceeding 20 ng/ml and levels of serum amyloid A exceeding 80 ng/ml. Double-stranded RNA, lipopolysaccharide, zymosan A, and flagellin also induced expression of macrophage inflammatory protein-3alpha and granulocyte-macrophage colony-stimulating factor, which may facilitate immature dendritic cell migration and maturation. These results suggest that airway epithelial cells express several TLRs and that they are functionally active. Epithelial expression of TLRs may be of importance in inflammation and immunity in the airways in response to inhaled pathogens.