Response of airway epithelial cells to double-stranded RNA in an allergic environment.
Response of airway epithelial cells to double-stranded RNA in an allergic environment.
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DOI：
10.1186/s40247-014-0011-6
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发表时间：
2014-12
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影响因子：
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通讯作者：
Kumar RK
中科院分区：
文献类型：
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作者：
Herbert C;Zeng QX;Shanmugasundaram R;Garthwaite L;Oliver BG;Kumar RK
呼吸道病毒感染是过敏性哮喘患者急性加重的最常见诱因。哮喘患者气道上皮细胞（AEC）的抗病毒反应可能受损，而AEC产生的细胞因子可能驱动炎症反应。我们研究了在与过敏性环境相关的Th 2细胞因子存在下培养的AEC是否对双链RNA（一种病毒样刺激）表现出改变的反应。我们进行了初步研究，使用MLE-12细胞系来自小鼠远端呼吸道上皮细胞，然后证实和扩展我们的研究结果，使用低传代人类AEC。将细胞在不存在或存在Th 2细胞因子IL-4和IL-13的情况下培养48小时，然后用聚I：C刺激4小时。通过定量实时PCR评估相关抗病毒应答和细胞因子基因的表达。通过免疫测定法评估细胞因子蛋白的分泌。在用poly I：C刺激后，用Th 2细胞因子预处理的MLE-12细胞表现出细胞因子基因Cxcl 10和Cxcl 11的mRNA表达水平显著更高，以及Cxcl 9和Il 6表达增加的趋势。抗病毒应答基因的表达大部分不变，尽管在Th 2细胞因子预处理的细胞中Stat 1、Ifit 1和Ifitm 3显著增加。用IL-4和IL-13预处理，然后用poly I：C刺激的人AEC类似地表现出显著更高的IL 8、CXCL 9、CXCL 10、CXCL 11和CCL 5基因表达。同时，CXCL 8和CCL 5的分泌显著增加，以及CXCL 10和IL-6的分泌增加的趋势。同样，抗病毒应答基因的表达没有降低。相反，III型干扰素、RNA解旋酶和其他干扰素刺激基因的mRNA表达显著增强。Th 2细胞因子环境似乎促进AEC响应于双链RNA而增加促炎趋化因子的产生，这可能有助于解释过敏性哮喘患者对呼吸道病毒感染的过度炎症反应。然而，哮喘患者抗病毒宿主防御的任何损害似乎不太可能是由AEC引起的Th 2型苏氨酸诱导的病毒应答基因表达下调的结果。本文的在线版本（doi：10.1186/s40247-014-0011-6）包含补充材料，可供授权用户使用。
Respiratory viral infections are the most common trigger of acute exacerbations in patients with allergic asthma. The anti-viral response of airway epithelial cells (AEC) may be impaired in asthmatics, while cytokines produced by AEC may drive the inflammatory response. We investigated whether AEC cultured in the presence of Th2 cytokines associated with an allergic environment exhibited altered responses to double-stranded RNA, a virus-like stimulus. We undertook preliminary studies using the MLE-12 cell line derived from mouse distal respiratory epithelial cells, then confirmed and extended our findings using low-passage human AEC. Cells were cultured in the absence or presence of the Th2 cytokines IL-4 and IL-13 for 48 hours, then stimulated with poly I:C for 4 hours. Expression of relevant anti-viral response and cytokine genes was assessed by quantitative real-time PCR. Secretion of cytokine proteins was assessed by immunoassay. Following stimulation with poly I:C, MLE-12 cells pre-treated with Th2 cytokines exhibited significantly higher levels of expression of mRNA for the cytokine genes Cxcl10 and Cxcl11, as well as a trend towards increased expression of Cxcl9 and Il6. Expression of anti-viral response genes was mostly unchanged, although Stat1, Ifit1 and Ifitm3 were significantly increased in Th2 cytokine pre-treated cells. Human AEC pre-treated with IL-4 and IL-13, then stimulated with poly I:C, similarly exhibited significantly higher expression of IL8, CXCL9, CXCL10, CXCL11 and CCL5 genes. In parallel, there was significantly increased secretion of CXCL8 and CCL5, as well as a trend towards increased secretion of CXCL10 and IL-6. Again, expression of anti-viral response genes was not decreased. Rather, there was significantly enhanced expression of mRNA for type III interferons, RNA helicases and other interferon-stimulated genes. The Th2 cytokine environment appears to promote increased production of pro-inflammatory chemokines by AEC in response to double-stranded RNA, which could help explain the exaggerated inflammatory response to respiratory viral infection in allergic asthmatics. However, any impairment of anti-viral host defences in asthmatics appears unlikely to be a consequence of Th2 cytokine-induced downregulation of the expression of viral response genes by AEC. The online version of this article (doi:10.1186/s40247-014-0011-6) contains supplementary material, which is available to authorized users.