鼻病毒持续存在的转录组学揭示体外鼻上皮细胞中RIG-I和干扰素刺激基因的持续表达
2022/09/20
摘要
背景：人类鼻病毒（HRVs）经常与哮喘恶化相关，并在哮喘患者的气道中发现。虽然HRV诱导的急性感染已有大量文献记载，但鼻上皮是否会持续HRV感染以及相关的宿主免疫反应激活尚不清楚。
目的：研究HRV持续期间人类鼻上皮细胞（hNECs）的持续调节宿主反应。
方法：通过时间过程研究，使用hNECs的体外感染模型建立了HRV16的持久性和病毒复制的动态。在感染后3天和14天（dpi），分别对感染早期和后期的hNECs进行RNA测序。使用基因本体论（GO）和Ingenuity通路分析对差异表达基因（DEGs）的功能富集进行了评估。
结果：HRV RNA和蛋白质的表达在整个长期感染过程中持续存在，即使在感染病毒子代产量减少后。对独特的DEGs的GO分析表明，在3dpi时与纤毛功能和气道重塑有关的途径的调节发生了改变，在14dpi时与丝氨酸型内肽酶活性有关。两个时间点之间共有的DEG的功能富集与干扰素（IFN）和细胞质模式识别受体（PRR）信号传导途径有关。对候选基因的持续调控的验证证实了RIG-I的持续表达，并揭示了它在HRV持续期间与干扰素刺激基因（ISG）的密切共调。
结论：在长期感染过程中，HRV RNA的持续存在并不一定表明有活动性感染。RIG-I和ISG在应对病毒RNA持续存在时的持续表达，突出了评估免疫激活宿主因素在活跃的HRV感染期间如何变化以及此后持续的免疫调节的重要性。
Transcriptomics of rhinovirus persistence reveals sustained expression of RIG-I and interferon-stimulated genes in nasal epithelial cells in vitro
Ong, H. H., Andiappan, A. K., Duan, K., Lum, J., Liu, J., Tan, K. S., Howland, S., Lee, B., Ong, Y. K., Thong, M., Chow, V. T., & Wang, D. Y.
Abstract
BACKGROUND:Human rhinoviruses (HRVs) are frequently associated with asthma exacerbations, and have been found in the airways of asthmatic patients. While HRV-induced acute infection is well-documented, it is less clear whether the nasal epithelium sustains prolonged HRV infections along with the associated activation of host immune responses.
OBJECTIVE:To investigate sustainably regulated host responses of human nasal epithelial cells (hNECs) during HRV persistence.
METHODS:Using a time-course study, HRV16 persistence and viral replication dynamics were established using an in vitro infection model of hNECs. RNA sequencing was performed on hNECs in the early and late stages of infection at 3 and 14 days post-infection (dpi), respectively. The functional enrichment of differentially expressed genes (DEGs) was evaluated using gene ontology (GO) and Ingenuity pathway analysis.
RESULTS:HRV RNA and protein expression persisted throughout prolonged infections, even after decreased production of infectious virus progeny. GO analysis of unique DEGs indicated altered regulation of pathways related to ciliary function and airway remodeling at 3 dpi and serine-type endopeptidase activity at 14 dpi. The functional enrichment of shared DEGs between the two time-points was related to interferon (IFN) and cytoplasmic pattern recognition receptor (PRR) signaling pathways. Validation of the sustained regulation of candidate genes confirmed the persistent expression of RIG-I and revealed its close co-regulation with interferon-stimulated genes (ISGs) during HRV persistence.
CONCLUSION:The persistence of HRV RNA does not necessarily indicate an active infection during prolonged infection. The sustained expression of RIG-I and ISGs in response to viral RNA persistence highlights the importance of assessing how immune-activating host factors can change during active HRV infection and the immune regulation that persists thereafter.
背景：人类鼻病毒（HRVs）经常与哮喘恶化相关，并在哮喘患者的气道中发现。虽然HRV诱导的急性感染已有大量文献记载，但鼻上皮是否会持续HRV感染以及相关的宿主免疫反应激活尚不清楚。
目的：研究HRV持续期间人类鼻上皮细胞（hNECs）的持续调节宿主反应。
方法：通过时间过程研究，使用hNECs的体外感染模型建立了HRV16的持久性和病毒复制的动态。在感染后3天和14天（dpi），分别对感染早期和后期的hNECs进行RNA测序。使用基因本体论（GO）和Ingenuity通路分析对差异表达基因（DEGs）的功能富集进行了评估。
结果：HRV RNA和蛋白质的表达在整个长期感染过程中持续存在，即使在感染病毒子代产量减少后。对独特的DEGs的GO分析表明，在3dpi时与纤毛功能和气道重塑有关的途径的调节发生了改变，在14dpi时与丝氨酸型内肽酶活性有关。两个时间点之间共有的DEG的功能富集与干扰素（IFN）和细胞质模式识别受体（PRR）信号传导途径有关。对候选基因的持续调控的验证证实了RIG-I的持续表达，并揭示了它在HRV持续期间与干扰素刺激基因（ISG）的密切共调。
结论：在长期感染过程中，HRV RNA的持续存在并不一定表明有活动性感染。RIG-I和ISG在应对病毒RNA持续存在时的持续表达，突出了评估免疫激活宿主因素在活跃的HRV感染期间如何变化以及此后持续的免疫调节的重要性。
(中日友好医院呼吸与危重症医学科 沈焜路 摘译 林江涛 审校)
(Allergy. 2022 Mar 16. DOI: 10.1111/all.15280)
(Allergy. 2022 Mar 16. DOI: 10.1111/all.15280)
Transcriptomics of rhinovirus persistence reveals sustained expression of RIG-I and interferon-stimulated genes in nasal epithelial cells in vitro
Ong, H. H., Andiappan, A. K., Duan, K., Lum, J., Liu, J., Tan, K. S., Howland, S., Lee, B., Ong, Y. K., Thong, M., Chow, V. T., & Wang, D. Y.
Abstract
BACKGROUND:Human rhinoviruses (HRVs) are frequently associated with asthma exacerbations, and have been found in the airways of asthmatic patients. While HRV-induced acute infection is well-documented, it is less clear whether the nasal epithelium sustains prolonged HRV infections along with the associated activation of host immune responses.
OBJECTIVE:To investigate sustainably regulated host responses of human nasal epithelial cells (hNECs) during HRV persistence.
METHODS:Using a time-course study, HRV16 persistence and viral replication dynamics were established using an in vitro infection model of hNECs. RNA sequencing was performed on hNECs in the early and late stages of infection at 3 and 14 days post-infection (dpi), respectively. The functional enrichment of differentially expressed genes (DEGs) was evaluated using gene ontology (GO) and Ingenuity pathway analysis.
RESULTS:HRV RNA and protein expression persisted throughout prolonged infections, even after decreased production of infectious virus progeny. GO analysis of unique DEGs indicated altered regulation of pathways related to ciliary function and airway remodeling at 3 dpi and serine-type endopeptidase activity at 14 dpi. The functional enrichment of shared DEGs between the two time-points was related to interferon (IFN) and cytoplasmic pattern recognition receptor (PRR) signaling pathways. Validation of the sustained regulation of candidate genes confirmed the persistent expression of RIG-I and revealed its close co-regulation with interferon-stimulated genes (ISGs) during HRV persistence.
CONCLUSION:The persistence of HRV RNA does not necessarily indicate an active infection during prolonged infection. The sustained expression of RIG-I and ISGs in response to viral RNA persistence highlights the importance of assessing how immune-activating host factors can change during active HRV infection and the immune regulation that persists thereafter.
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