Background

Cystic fibrosis (CF), a genetic disease caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, is characterized by dysfunction of the immune response in the airway epithelium that leads to prolonged infection, colonization and exacerbated inflammation. In this study, we determined the gene expression profile of airway epithelial cells knockdown for CFTR (CFTR KD) in response to bacterial and viral challenges.

Methods

In a first approach, polarized CFTR KD and their control counterpart (CFTR CTL) cells were stimulated with P. aeruginosa-derived virulence factor flagellin. Next, we developed a model of Influenza A virus (IAV) infection in CTL and CFTR KD polarized cells. mRNA was collected for transcriptome analysis.

Results

Beside the expected pro-inflammatory response, Gene Set Enrichment Analysis highlighted key molecular pathways and players involved in IAV and anti-viral interferon signaling. Although IAV replication was similar in both cell types, multiplex gene expression analysis revealed changes of key immune genes dependent on time of infection that were found to be CFTR-dependent and/or IAV-dependent. Interferons are key signaling proteins/cytokines in the antibacterial and antiviral response. To evaluate their impact on the altered gene expression profile in CFTR responses to pathogens, we measured transcriptome changes after exposure to Type I-, Type II- and Type III-interferons.

Conclusions

Our findings reveal target genes in understanding the defective immune response in the CF airway epithelium in the context of viral infection. Information provided in this study would be useful to understand the dysfunctional immune response of the CF airway epithelium during infection.

中文翻译：

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感染甲型流感病毒的极化囊性纤维化气道上皮细胞的免疫反应

背景

囊性纤维化 (CF) 是一种由囊性纤维化跨膜电导调节因子 ( CFTR ) 基因突变引起的遗传性疾病，其特征是气道上皮细胞免疫反应功能障碍，导致感染延长、定植和炎症加剧。在这项研究中，我们确定了针对细菌和病毒挑战的 CFTR (CFTR KD) 气道上皮细胞敲低的基因表达谱。

方法

在第一种方法中，极化的 CFTR KD 和它们的对照对应物 (CFTR CTL) 细胞用铜绿假单胞菌衍生的毒力因子鞭毛蛋白刺激。接下来，我们在 CTL 和 CFTR KD 极化细胞中开发了甲型流感病毒 (IAV) 感染模型。收集mRNA用于转录组分析。

结果

除了预期的促炎反应外，基因集富集分析还强调了参与 IAV 和抗病毒干扰素信号传导的关键分子途径和参与者。尽管 IAV 复制在两种细胞类型中相似，但多重基因表达分析揭示了依赖于感染时间的关键免疫基因的变化，这些基因被发现是 CFTR 依赖性和/或 IAV 依赖性的。干扰素是抗菌和抗病毒反应中的关键信号蛋白/细胞因子。为了评估它们对 CFTR 对病原体反应中基因表达谱改变的影响，我们测量了暴露于 I 型、II 型和 III 型干扰素后的转录组变化。

结论

我们的研究结果揭示了了解病毒感染情况下 CF 气道上皮细胞中缺陷免疫反应的靶基因。本研究中提供的信息将有助于了解 CF 气道上皮在感染期间的功能失调免疫反应。