Cellular crosstalk between airway epithelial and endothelial cells regulates barrier functions during exposure to double-stranded RNA.
Cellular crosstalk between airway epithelial and endothelial cells regulates barrier functions during exposure to double-stranded RNA.
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DOI：
10.1002/iid3.139
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发表时间：
2017-03
期刊：
影响因子：
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通讯作者：
Davies DE
中科院分区：
文献类型：
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作者：
Blume C;Reale R;Held M;Loxham M;Millar TM;Collins JE;Swindle EJ;Morgan H;Davies DE
气道粘膜的上皮和内皮屏障对于调节组织稳态和保护免受病原体或其他组织损伤剂的影响至关重要。为了应对病毒感染，上皮细胞必须向内皮发出信号以启动免疫细胞募集。这是一个高度时间调节的过程;然而，这种串扰的机制尚未完全理解。在人气道上皮和内皮细胞的紧密接触共培养模型中，使用跨上皮电阻（TER）测量、免疫荧光、电子显微镜和ELISA分析细胞串扰。通过将气道上皮细胞顶部暴露于双链RNA（Poly（I：C））来模拟病毒感染。使用微流控培养系统，在共培养模型中分析了介体的暂时释放。在激发后4小时内，双链RNA诱导上皮细胞释放TNF-α。这通过激发后8小时触发化学引诱物CX 3CL 1（fractalkine）的释放以及粘附分子E-选择素和ICAM-1的表达来激活内皮细胞。中和TNF-α可显著降低这些反应。通过促进动力学分析，微流体共培养系统已经能够识别上皮和内皮屏障之间的关键信号传导机制。更好地了解细胞间串扰及其调节机制有可能确定新的治疗策略来控制气道炎症。
The epithelial and endothelial barriers of the airway mucosa are critical for regulation of tissue homeostasis and protection against pathogens or other tissue damaging agents. In response to a viral infection, epithelial cells must signal to the endothelium to initiate immune cell recruitment. This is a highly temporal regulated process; however, the mechanisms of this cross‐talk are not fully understood. In a close‐contact co‐culture model of human airway epithelial and endothelial cells, cellular crosstalk was analyzed using transepithelial electrical resistance (TER) measurements, immunofluorescence, electron microscopy, and ELISA. Viral infections were simulated by exposing airway epithelial cells apically to double‐stranded RNA (Poly(I:C)). Using a microfluidic culture system, the temporal release of mediators was analyzed in the co‐culture model. Within 4 h of challenge, double‐stranded RNA induced the release of TNF‐α by epithelial cells. This activated endothelial cells by triggering the release of the chemoattractant CX3CL1 (fractalkine) by 8 h post‐challenge and expression of adhesion molecules E‐selectin and ICAM‐1. These responses were significantly reduced by neutralising TNF‐α. By facilitating kinetic profiling, the microfluidic co‐culture system has enabled identification of a key signaling mechanism between the epithelial and endothelial barriers. Better understanding of cell–cell cross‐talk and its regulatory mechanisms has the potential to identify new therapeutic strategies to control airway inflammation.