人冠状病毒是重要的呼吸道感染病原，其中HCoV-HKU1可引起人重症肺炎。因常规细胞培养技术难于获得HCoV-HKU1分离株，国际上对不同HCoV-HKU1临床感染毒株的复制及生物学特性的研究甚少。分化良好的人呼吸道上皮细胞（HAE）是冠状病毒的天然宿主细胞，也是目前世界上唯一能够分离培养HCoV-HKU1的细胞。针对我国尚无HCoV-HKU1毒株和缺乏HCoV-HKU1生物学特性研究的现状，本课题组将前期已经建立的HAE培养体系应用于急性呼吸道感染（特别是重症肺炎）患者中HCoV-HKU1的分离鉴定，获得了一株HCoV-HKU1临床分离株。我们拟将进一步扩大临床样本中HCoV-HKU1的分离工作，研究 HCoV-HKU1在HAE模型中的复制特点；通过序列分析研究其遗传变异规律；并通过细胞生物学研究方法，阐明其亚细胞定位和细胞内转运等生物学特性，为人冠状病毒致病性研究及科学诊治奠定基础。

As one of the most important pathogens of humans, human coronavirus HKU1 (HCoV-HKU1) can cause acute respiratory tract illness, especially in Children. But little is known about the replication and characterization of HCoV-HKU1, as all attempts to culture a clinical isolate of HCoV-HKU1 have failed in standard transformed cell cultures. Well differentiated human airway epithelial cell culture system (HAE) is an vitro model of the human airway epithelium. HAE recapitulate the morphological and physiological features of the airway epithelium in vivo and is comprised of ciliated, non-ciliacted mucin-secreting and basal epithelial cells, and it is the only one cell culture system to successfully propagate HCoV-HKU1 in vitro.As there is no HCoV-HKU1 stain isolated in China and little is known about the characterization of HCoV-HKU1 in China, we use HAE system, which has been established in our lab, to culture the clinical isolates of HCoV-HKU1.Now we have gained one strain of HCoV-HKU1. We will continue the isolation of HCoV-HKU1 from clinical material to get more stains of HCoV-HKU1 in China. Meanwhile, we will get the complete genome sequence of HCoV-HKU1 stains and subsequent passage HCoV-HKU1 stains in HAE to know about the basic characterization of HCoV-HKU1, such as the replication kinetics, the cellular localization and so on.Our work will provide basic data about pathogenisis and diagnosis of human coronaviruses.