Rhinovirus upregulates transient receptor potential channels in a human neuronal cell line: implications for respiratory virus-induced cough reflex sensitivity
Rhinovirus upregulates transient receptor potential channels in a human neuronal cell line: implications for respiratory virus-induced cough reflex sensitivity
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DOI：
10.1136/thoraxjnl-2013-203894
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发表时间：
2014-01-01
期刊：
影响因子：
10
通讯作者：
McGarvey, L. P. A.
中科院分区：
文献类型：
--
作者：
Abdullah, H.;Heaney, L. G.;McGarvey, L. P. A.
背景呼吸道病毒引起咳嗽过敏的机制尚不清楚。负责感知物理和化学刺激的气道神经元受体的上调是一种可能性，并且瞬时受体电位（TRP）通道家族是潜在的候选者。我们已经使用了在体外模型的感觉神经元和人鼻病毒（HRV-16）研究病毒感染对TRP expression.Methods IMR-32神经母细胞瘤细胞在培养中分化表达三个TRP通道：TRPV 1，TRPA 1和TRPM 8的影响。使用流式细胞术和qRT-PCR测量接种活病毒、灭活病毒、病毒诱导的可溶性因子或沉淀病毒颗粒后的TRP通道蛋白和mRNA水平。结果感染后2-4 h，TRPA 1和TRPV 1表达均出现早期上调，而感染后2-4 h，TRPA 1和TRPV 1表达均出现早期上调。这与复制病毒无关，因为单独的病毒诱导的可溶性因子足以分别使通道表达增加50倍和15倍。感染细胞上清液中增加的NGF、IL-6和IL-8水平代表可能的候选物。与此相反，TRPM 8的表达是最大的48小时（9.6倍），需要病毒复制，而不是可溶性factors.Conclusions我们首次表明，鼻病毒可以感染神经细胞。此外，感染通过通道特异性机制引起TRP通道的上调。TRPA 1和TRPV 1水平的增加可以由感染诱导的可溶性因子介导，而TRPM 8需要复制病毒。TRP通道可能是控制病毒引起的咳嗽的新的治疗靶点。
Background The mechanism underlying respiratory virus-induced cough hypersensitivity is unknown. Upregulation of airway neuronal receptors responsible for sensing physical and chemical stimuli is one possibility, and the transient receptor potential (TRP) channel family are potential candidates. We have used an in vitro model of sensory neurons and human rhinovirus (HRV-16) to study the effect of virus infection on TRP expression.Methods IMR-32 neuroblastoma cells were differentiated in culture to express three TRP channels: TRPV1, TRPA1 and TRPM8. Flow cytometry and qRT-PCR were used to measure TRP channel protein and mRNA levels following inoculation with live virus, inactivated virus, virus-induced soluble factors or pelleted virus particles. Multiplex bioassay was used to determine nerve growth factor (NGF), interleukin (IL)-1 beta, IL-6 and IL-8 levels in response to infection.Results Early upregulation of TRPA1 and TRPV1 expression occurred 2-4 h post infection. This was independent of replicating virus as virus-induced soluble factors alone were sufficient to increase channel expression 50-fold and 15-fold, respectively. NGF, IL-6 and IL-8 levels, increased in infected cell supernatants, represent possible candidates. In contrast, TRPM8 expression was maximal at 48 h (9.6-fold) and required virus replication rather than soluble factors.Conclusions We show for the first time that rhinovirus can infect neuronal cells. Furthermore, infection causes upregulation of TRP channels by channel-specific mechanisms. The increase in TRPA1 and TRPV1 levels can be mediated by soluble factors induced by infection whereas TRPM8 requires replicating virus. TRP channels may be novel therapeutic targets for controlling virus-induced cough.