TNF-α induces vectorial secretion of IL-8 in Caco-2 cells.
TNF-α induces vectorial secretion of IL-8 in Caco-2 cells.
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DOI：
10.1007/s11605-010-1321-9
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发表时间：
2010-10
期刊：
影响因子：
--
通讯作者：
Pritts TA
中科院分区：
文献类型：
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作者：
Sonnier DI;Bailey SR;Schuster RM;Lentsch AB;Pritts TA
肠上皮细胞是先天免疫的重要组成部分，对炎症刺激具有复杂的反应。肠上皮细胞极性影响炎症刺激反应的方式在很大程度上是未知的。我们假设极化肠上皮细胞表现出依赖于刺激位置的双向炎症反应。Caco-2细胞在双室培养系统的半透性插入物上生长，并在根尖室或基底外室用肿瘤坏死因子-α (TNF-α; 100 ng/ml)或无血清培养基处理。采用酶联免疫吸附法测定各组白细胞介素-8 （IL-8）的产生。为了确定受体特异性，将抗tnf受体抗体添加到根尖室或基底外侧室。基底外侧刺激TNF-α可增加根尖和基底外侧IL-8的产生。根尖TNF-α刺激导致根尖分泌增加，但基底外侧无IL-8分泌。受体阻断提示TNF受体1参与根尖膜和基底膜，而TNF受体2仅参与根尖膜。极化肠上皮细胞对TNF-α刺激的反应是集中、定向地分泌促炎细胞因子IL-8。这些发现很重要，因为它们表明肠上皮细胞能够组织对炎症信号的反应，并以双向的载体方式产生炎症介质。
Intestinal epithelial cells represent an important component of innate immunity, with sophisticated responses to inflammatory stimuli. The manner in which intestinal epithelial cell polarity affects responses to inflammatory stimuli is largely unknown. We hypothesized that polarized intestinal epithelial cells exhibit a bidirectional inflammatory response dependent upon the location of the stimulus. Caco-2 cells were grown on semi-permeable inserts in a dual-compartment culture system and treated with tumor necrosis factor-α (TNF-α; 100 ng/ml) or serum-free media in the apical or basolateral chamber. Interleukin-8 (IL-8) production in each chamber was measured by enzyme-linked immunosorbent assay. To determine receptor specificity, anti-TNF receptor antibodies were added to the apical or basolateral chamber. Basolateral stimulation with TNF-α resulted in increased apical and basolateral IL-8 production. Apical TNF-α stimulation resulted in increased apical, but not basolateral IL-8 production. Receptor blockade suggested TNF receptor 1 involvement on both apical and basolateral membranes, while TNF receptor 2 was only active on the apical membrane. Polarized intestinal epithelial cells respond to TNF-α stimulation with focused, directional secretion of the proinflammatory cytokine IL-8. These findings are important because they suggest that intestinal epithelial cells are capable of organizing their response to inflammatory signals and producing inflammatory mediators in a bidirectional, vectorial fashion.