Center for Immunity and Immunotherapies, Seattle Children's Research Institute, 1900 Ninth Ave., Seattle, WA, 98145, USA.
Department of Genome Sciences, University of Washington, Seattle, WA, USA.
Sci Rep. 2022 Apr 28;12(1):6972. doi: 10.1038/s41598-022-10763-2.
Common alphacoronaviruses and human rhinoviruses (HRV) induce type I and III interferon (IFN) responses important to limiting viral replication in the airway epithelium. In contrast, highly pathogenic betacoronaviruses including SARS-CoV-2 may evade or antagonize RNA-induced IFN I/III responses. In airway epithelial cells (AECs) from children and older adults we compared IFN I/III responses to SARS-CoV-2 and HRV-16, and assessed whether pre-infection with HRV-16, or pretreatment with recombinant IFN-β or IFN-λ, modified SARS-CoV-2 replication. Bronchial AECs from children (ages 6-18 years) and older adults (ages 60-75 years) were differentiated ex vivo to generate organotypic cultures. In a biosafety level 3 (BSL-3) facility, cultures were infected with SARS-CoV-2 or HRV-16, and RNA and protein was harvested from cell lysates 96 h. following infection and supernatant was collected 48 and 96 h. following infection. In additional experiments cultures were pre-infected with HRV-16, or pre-treated with recombinant IFN-β1 or IFN-λ2 before SARS-CoV-2 infection. In a subset of experiments a range of infectious concentrations of HRV-16, SARS-CoV-2 WA-01, SARS-CoV-2 Delta variant, and SARS-CoV-2 Omicron variant were studied. Despite significant between-donor heterogeneity SARS-CoV-2 replicated 100 times more efficiently than HRV-16. IFNB1, INFL2, and CXCL10 gene expression and protein production following HRV-16 infection was significantly greater than following SARS-CoV-2. IFN gene expression and protein production were inversely correlated with SARS-CoV-2 replication. Treatment of cultures with recombinant IFNβ1 or IFNλ2, or pre-infection of cultures with HRV-16, markedly reduced SARS-CoV-2 replication. In addition to marked between-donor heterogeneity in IFN responses and viral replication, SARS-CoV-2 (WA-01, Delta, and Omicron variants) elicits a less robust IFN response in primary AEC cultures than does rhinovirus, and heterologous rhinovirus infection, or treatment with recombinant IFN-β1 or IFN-λ2, reduces SARS-CoV-2 replication, although to a lesser degree for the Delta and Omicron variants.
常见的α冠状病毒和人类鼻病毒（HRV）诱导 I 型和 III 型干扰素（IFN）反应，对限制气道上皮细胞中的病毒复制很重要。相比之下，高致病性β冠状病毒，包括 SARS-CoV-2，可能逃避或拮抗 RNA 诱导的 IFN I/III 反应。我们比较了 SARS-CoV-2 和 HRV-16 在儿童和老年人群的气道上皮细胞（AEC）中的 IFN I/III 反应，并评估了 HRV-16 预先感染或重组 IFN-β或 IFN-λ 预处理是否改变了 SARS-CoV-2 的复制。从儿童（6-18 岁）和老年人（60-75 岁）中分离出支气管 AEC 并在体外分化为器官型培养物。在生物安全级别 3（BSL-3）设施中，用 SARS-CoV-2 或 HRV-16 感染培养物，并在感染后 96 小时从细胞裂解物中提取 RNA 和蛋白质，并在感染后 48 和 96 小时收集上清液。在其他实验中，在感染 SARS-CoV-2 之前，培养物预先感染 HRV-16 或用重组 IFN-β1 或 IFN-λ2 预处理。在一部分实验中，研究了一系列感染性浓度的 HRV-16、SARS-CoV-2 WA-01、SARS-CoV-2 Delta 变体和 SARS-CoV-2 Omicron 变体。尽管存在显著的供体间异质性，但 SARS-CoV-2 的复制效率比 HRV-16 高 100 倍。HRV-16 感染后 IFNB1、INFL2 和 CXCL10 基因表达和蛋白产生显著高于 SARS-CoV-2。IFN 基因表达和蛋白产生与 SARS-CoV-2 复制呈负相关。用重组 IFNβ1 或 IFNλ2 处理培养物或用 HRV-16 预先感染培养物，均可显著降低 SARS-CoV-2 的复制。除了 IFN 反应和病毒复制的显著供体间异质性外，SARS-CoV-2（WA-01、Delta 和 Omicron 变体）在原代 AEC 培养物中引起的 IFN 反应比鼻病毒弱，异源鼻病毒感染或用重组 IFN-β1 或 IFN-λ2 处理，虽然对 Delta 和 Omicron 变体的效果要小一些，但也能降低 SARS-CoV-2 的复制。
