抗ALV-J的鸡干扰素刺激基因鉴定及其抗病毒机制研究

As host antiviral genes, interferon stimulated genes (ISGs) have high potential to be used as antiviral candidate genes for breeding of disease resistance. However, the identified chicken ISGs were limited and the relevant antiviral molecular mechanism of chicken ISGs is rarely involved. In this study, chicken type I, II, III ISGs and ISGs with anti-ALV-J potential will be primarily identified by RNA-Seq. Based on of this result, chicken ISGs against ALV-J replication will be confirmed using over-expression and RNAi. Their sequence variations in Chinese local chicken breeds will be analyzed by target region capture sequencing technology. Finally, in order to elucidate the anti-ALV-J molecular mechanism of chicken ISGs, the interaction between chicken ISGs and host factor or virus protein will be analyzed and the effect of chicken ISGs on the immune pathway will be studied by pull-down, Co-IP, and etc. The results of this study will provide valuable basic materials for ALV-J resistant breeding. It has important implications for breeding disease-resistant layers and broilers.

干扰素刺激基因(ISGs)作为宿主抗病毒基因，是极具潜力的抗病育种候选基因。目前已鉴定出来的具有抗病毒活性的鸡ISGs还很有限，相关的抗病毒分子机制研究更是鲜有涉及。本项目拟首先通过RNA-Seq系统鉴定出鸡I、II、III型ISGs以及潜在的抗J亚群禽白血病病毒(ALV-J)的鸡ISGs；然后利用过表达和RNA干扰等方法确认对ALV-J具有抗性的ISGs；再通过目标区域捕获测序技术，分析这些抗ALV-J的鸡ISGs在我国地方鸡种中的序列变异；最后通过蛋白pull-down、Co-IP等方法，分析抗ALV-J的鸡ISGs与宿主因子或病毒蛋白的互作关系，明确其对宿主抗病毒免疫通路的影响，阐明鸡ISGs抗ALV-J的分子机制。这将为抗ALV-J育种研究提供宝贵的基础材料，对培育抗ALV-J的鸡品种有重要意义。

干扰素刺激基因（ISGs）是宿主体内一群数量庞大的抗病毒基因，几乎可以靶向病毒复制周期的每一个环节。阻断病毒生命周期的任一环节，均可达到抗病毒的效果。目前在家禽中，只有Mx、IFITM3和ZAP等少数几个经典ISGs 被鉴定出来，并被证明具有抗病毒功能。还有大量的家鸡 ISGs 有待被鉴定及进一步的抗病毒机制研究。J 亚群禽白血病病毒（ALV-J）是一种诱发鸡发生免疫抑制及产生肿瘤的逆转录病毒，既能水平传播又能垂直传播，给我国养禽业造成了巨大的损失。本项目系统鉴定出了家鸡的ISGs并阐明了其中个别抗ALV-J ISG的作用机制。通过RNA-Seq，我们在PBMC、DF1以及LMH等家禽细胞上系统鉴定出鸡I、II、III型干扰素诱导产生的ISGs数百个。通过多种种体外验证实验阐明了CH25H通过产生25HC抑制ALV-J的入侵以及CH25H通过与CHMP4B 的互作调控宿主细胞的自噬来抑制ALV-J复制的机制；EAF2主要利用细胞代谢通路来抑制ALV-J的复制的机制。本项目的实施为家鸡ISGs的研究提供了宝贵素材，为家鸡抗病毒育种研究提供宝贵的基础材料。