HSP47蛋白与禽传染性支气管炎病毒（IBV）SAIBK株感染宿主细胞的相关性研究

IBV are divided into five tpyes according to the histotropism of the virus, such as respiratory type, proventriculus type, kidney type, intestinal type, and genital tract type. The IBV of kidney type is highly infectious and prevalent in China, which is serious damage to poultry industry. The SAIBK strain is kidney type of IBV, and we have proved that HSP47 could be interact with S1 protein of the strain in chicken. The scientific question is that whether HSP47 has the correlation of the infection of host cells of IBV. The study content includes the following two points. Firstly, constrcting the real-time PCR method for HSP47, and studying differential expression of the HSP47 mRNA in different tissues and organs in SPF chicken after infecting SAIBK strain, in order to get the relationship between HSP47 and the contents of IBV in different infecting time and tissues, and to analyze whether HSP47 has the correlation of the histotropism of IBV. Secondly, using RNAi to interfere HSP47 genes' expression, to study the relationship between expression level of HSP47 and the virus infection of cells. The study provides not only scientific basis for revealing the correlation between HSP47 and the IBV SAIBK strain infecting host cells, but also new ideas for the prevention and control of IBV.

禽传染性支气管炎病毒（IBV）根据对组织的亲嗜性可分为呼吸型、腺胃型、肾型、肠型、生殖道型五个类型，其中肾型传染性强、流行范围广，对养禽业危害严重。SAIBK株为IBV肾型毒株，申请者已证实鸡体内的热休克蛋白47（HSP47）能够与IBV SAIBK株的S1蛋白相互作用。科学问题是,HSP47是否与IBV感染宿主细胞有关？拟开展以下研究：建立检测HSP47的相对定量荧光PCR方法，对IBV SAIBK株感染SPF鸡后不同组织器官HSP47 mRNA的差异表达进行研究，掌握HSP47与不同时间段、不同组织IBV感染量的关系，分析HSP47是否与病毒的组织嗜性有关；应用RNAi技术干扰HSP47 mRNA表达研究其是否能抑制IBV对细胞的感染，掌握不同表达量的HSP47与IBV感染细胞的关系。以上研究对揭示HSP47与IBV感染宿主细胞的相关性提供了科学依据，也为IBV的防控提供了新的思路。

禽传染性支气管炎是由禽传染性支气管炎病毒病毒引起的危害全球养禽业的重要传染病之一。IBV血清型众多，不同血清型在细胞适应性上具有明显的差异。病毒-宿主蛋白的相互作用是目前研究的热点之一，诸多研究表明病毒通过与宿主细胞相互作用创造复制的条件。本项目主要开展了以下两项研究工作：1）通过荧光定量PCR方法检测了IBV不同毒株感染鸡胚原代肾细胞和鸡胚原代成纤维细胞后病毒的动态变化，同时建立鸡HSP47 mRNA荧光定量PCR方法并探究了在感染IBV病毒后细胞中HSP47 mRNA的差异表达，研究发现：在CEK、CEF细胞中HSP47的mRNA转录量与病毒含量成正相关，为研究HSP47mRNA的差异表达和病毒增殖量的相关性奠定基础。2）构建HSP47基因 CRISPR/Cas9 敲除体系，获得高效、稳定、永久性HSP47 敲除的非洲绿猴肾细胞株（Vero），Western blot测定Vero细胞中HSP47蛋白的表达量，IBV感染敲出细胞后其感染效率显著降低，说明鸡HSP47基因与IBV感染宿主细胞有关。以上研究对揭示HSP47对IBV感染宿主细胞的相关性提供了科学依据，同时也为IBV的防控提供了新的思路。