背景:慢性炎症性肺病急性加重,如慢性阻塞性肺疾病(COPD),通常与鼻病毒(RV)感染有关。尽管有这些协会,病毒引起的急性加重的发病机制尚不完全清楚.我们的目的是调查香烟烟雾(CS)的影响,COPD的主要危险因素,研究呼吸道上皮RV感染,并确定与这些作用相关的新机制。
方法:通过在气-液界面(ALI)培养并暴露于CS和RV-A16来分化来自COPD患者和对照组的原代支气管上皮细胞(PBEC)。使用在感染后6和24小时(hpi)收集的样品进行大量RNA测序,和病毒载量,在6、24和48hpi测量介质和L-乳酸水平。为了进一步描述CS对RV-A16感染的影响,我们进行了生长分化因子15(GDF15)敲除,ALI-PBEC中的L-乳酸和干扰素预处理。我们进行了去卷积分析,以预测各种暴露后ALI-PBEC细胞组成的变化。最后,我们比较了人RV-A16攻击后ALI-PBEC与鼻上皮的转录反应。
结果:在ALI-PBEC中,CS暴露在6hpi时损害了抗病毒反应,在24和48hpi时增加了病毒复制。在24hpi,CS暴露增强RV-A16诱导的上皮干扰素的表达,炎症相关基因和CXCL8。CS暴露增加氧化应激相关基因的表达,GDF15,线粒体膜电位降低。GDF15敲低实验表明该途径参与CS诱导的病毒复制增加。CS暴露增加糖酵解相关基因的表达和L-乳酸的产生,并被证明有助于更高的病毒复制。在COPD和非COPD来源的培养物之间没有发现重大差异。然而,细胞去卷积分析预测了基线时COPD源性培养物中较高的分泌细胞.
结论:总而言之,我们的研究结果表明,CS暴露导致更高的病毒感染在人支气管上皮通过改变不仅干扰素反应,但也可能通过转向糖酵解,并通过GDF15相关途径。
BACKGROUND: Acute exacerbations of chronic inflammatory lung diseases, such as chronic obstructive pulmonary disease (COPD), are frequently associated with rhinovirus (RV) infections. Despite these associations, the pathogenesis of virus-induced exacerbations is incompletely understood. We aimed to investigate effects of cigarette smoke (CS), a primary risk factor for COPD, on RV infection in airway epithelium and identify novel mechanisms related to these effects.
METHODS: Primary bronchial epithelial cells (PBEC) from COPD patients and controls were differentiated by culture at the air-liquid interface (ALI) and exposed to CS and RV-A16. Bulk RNA sequencing was performed using samples collected at 6 and 24 h post infection (hpi), and viral load, mediator and L-lactate levels were measured at 6, 24 and 48hpi. To further delineate the effect of CS on RV-A16 infection, we performed growth differentiation factor 15 (GDF15) knockdown, L-lactate and interferon pre-treatment in ALI-PBEC. We performed deconvolution analysis to predict changes in the cell composition of ALI-PBEC after the various exposures. Finally, we compared transcriptional responses of ALI-PBEC to those in nasal epithelium after human RV-A16 challenge.
RESULTS: CS exposure impaired antiviral responses at 6hpi and increased viral replication at 24 and 48hpi in ALI-PBEC. At 24hpi, CS exposure enhanced expression of RV-A16-induced epithelial interferons, inflammation-related genes and CXCL8. CS exposure increased expression of oxidative stress-related genes, of GDF15, and decreased mitochondrial membrane potential. GDF15 knockdown experiments suggested involvement of this pathway in the CS-induced increase in viral replication. Expression of glycolysis-related genes and L-lactate production were increased by CS exposure, and was demonstrated to contribute to higher viral replication. No major differences were demonstrated between COPD and non-COPD-derived cultures. However, cellular deconvolution analysis predicted higher secretory cells in COPD-derived cultures at baseline.
CONCLUSIONS: Altogether, our findings demonstrate that CS exposure leads to higher viral infection in human bronchial epithelium by altering not only interferon responses, but likely also through a switch to glycolysis, and via GDF15-related pathways.