Volatile emanations from in vitro airway cells infected with human rhinovirus
Volatile emanations from in vitro airway cells infected with human rhinovirus
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DOI：
10.1088/1752-7155/8/3/037110
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发表时间：
2014-09-01
影响因子：
3.8
通讯作者：
Davis, Cristina E.
中科院分区：
文献类型：
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作者：
Schivo, Michael;Aksenov, Alexander A.;Davis, Cristina E.
呼吸道病毒感染，如人类鼻病毒(HRV)，可导致相当大的发病率和死亡率，特别是在患有哮喘和慢性阻塞性肺病等潜在肺部疾病的人中。一种被提议的非侵入性检测病毒感染的策略是通过分析呼气来评估挥发性有机化合物(VOC)。上皮细胞是呼吸道感染过程中最重要的细胞系之一，是病原体防御的第一道防线。通过了解呼吸道上皮细胞挥发性有机碳的释放，可以极大地帮助发现感染特异性生物标志物。在这里，我们测试了一种假设，即从呼吸道细胞培养的顶腔获得的VOCs将区分健康细胞和感染HRV的细胞。原代培养的人气管、支气管细胞被放置在一个设计成捕获顶腔VOCs的系统中。将感染HRV的细胞与未感染的对照细胞进行比较。此外，用热灭活的HRV和TLR3激动剂Poly(I：C)处理的细胞与对照组进行比较。用固相微萃取纤维对顶空进行采样，用气相色谱/质谱仪分析挥发性有机物。我们确定了受感染细胞对脂肪醇、支链碳氢化合物和二甲基硫化物等化合物的差异表达，VOCs以前与氧化应激和细菌感染有关。我们没有发现致死的HRV、PolyI：C和对照细胞VOCs之间的主要差异。我们推测这些化合物可以作为HRV感染的生物标志物，VOCs的产生不是由于TLR3的刺激，而是需要主动的病毒复制。我们的新方法可以用于其他重要呼吸道病毒的体外研究，最终可能有助于识别病毒感染的VOC生物标记物，用于医疗诊断。
Respiratory viral infections such as human rhinovirus (HRV) can lead to substantial morbidity and mortality, especially in people with underlying lung diseases such as asthma and COPD. One proposed strategy to detect viral infections non-invasively is by volatile organic compound (VOC) assessment via analysis of exhaled breath. The epithelial cells are one of the most important cell lines affected during respiratory infections as they are the first line of pathogen defense. Efforts to discover infection-specific biomarkers can be significantly aided by understanding the VOC emanations of respiratory epithelial cells. Here we test the hypothesis that VOCs obtained from the headspace of respiratory cell culture will differentiate healthy cells from those infected with HRV. Primary human tracheobronchial cells were cultured and placed in a system designed to trap headspace VOCs. HRV-infected cells were compared to uninfected control cells. In addition, cells treated with heat-killed HRV and poly(I:C), a TLR3 agonist, were compared to controls. The headspace was sampled with solid-phase microextraction fibers and VOCs were analyzed by gas chromatography/mass spectrometry. We determined differential expression of compounds such as aliphatic alcohols, branched hydrocarbons, and dimethyl sulfide by the infected cells, VOCs previously associated with oxidative stress and bacterial infection. We saw no major differences between the killed-HRV, poly(I:C), and control cell VOCs. We postulate that these compounds may serve as biomarkers of HRV infection, and that the production of VOCs is not due to TLR3 stimulation but does require active viral replication. Our novel approach may be used for the in vitro study of other important respiratory viruses, and ultimately it may aid in identifying VOC biomarkers of viral infection for point-of-care diagnostics.