新城疫病毒Mukteswar株感染鸡巨噬细胞入胞及胞内转运途径研究  

Study on the Entry and Intracellular Transport Pathway of Newcastle Disease Virus Mukteswar Strain in Infected Chicken Macrophages

作　　者：黄新月 谭磊[2] 宋翠萍[2] 仇旭升[2] 孙英杰[2] 刘炜玮 廖瑛[2] 丁铲[2] 吴艳涛[1] HUANG Xinyue;TAN Lei;SONG Cuiping;QIU Xusheng;SUN Yingjie;LIU Weiwei;LIAO Ying;DING Chan;WU Yantao(College of Veterinary Medicine,Yangzhou University,Yangzhou 225000,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)

机构地区：[1]扬州大学兽医学院,扬州225000 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2024年第5期1-10,共10页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金面上项目(32072868);上海市自然科学基金面上项目(20ZR1469400);上海市兽医生物技术重点实验室开放基金资助(shklab202007)。

摘　　要：新城疫病毒(NDV)可以通过多种内吞途径入侵不同宿主细胞并在内体中逃逸从而避免被宿主天然免疫系统“追踪”。为探究NDV中等毒力疫苗株Mukteswar感染鸡巨噬细胞(HD11)的入胞和胞内转运途径,本研究利用Dynasore、CPZ、MβCD、Baf A1预处理细胞,然后感染NDV Mukteswar株,通过Westem blot和IFA检测NDV NP蛋白的表达变化。结果显示Dynasore、MβCD和Baf A1处理后NP的表达水平降低,表明NDV入侵HD11细胞是利用小窝蛋白介导的内吞途径,且依赖动力蛋白、胆固醇和pH。通过过表达和干扰试验检测调节不同内体囊泡类型运输的Rab蛋白(Rab5、Rab7、Rab11)对NDV感染HD11细胞的影响,结果发现Rab7的表达对NDV感染无影响,而过表达Rab5和Rab11促进NDV感染,表明是由Rab5、Rab11而不是Rab7调节NDV的转运。本研究丰富了NDV入侵宿主免疫细胞的途径及胞内转运的研究,并为全面阐明NDV感染机制提供了理论基础。Newcastle disease virus(NDV)can infect diff erent host cells through a variety of endocytosis pathways and escape from endosomes so as to avoid being tracked by the host’s innate immune system.In order to explore the pathways of entry and intracellular transport of mesogenic NDV vaccine strain Mukteswar in infected chicken macrophages(HD11),cells were pretreated with Dynasore,CPZ,MβCD and Baf A1 and then infected with NDV Mukteswar in this study.The changes of NDV NP protein expression were examined by Western blot and IFA.The results showed that the expression level of NP decreased after Dynasore,MβCD,Baf A1 treatment,indicating that NDV infected HD11 cells through the caveolae-mediated endocytosis depending on dynamin,cholesterol and pH.Subsequently,the eff ects of Rab proteins(Rab5,Rab7and Rab11)that regulated the transport of diff erent endosomal vesicle on NDV-infected HD11 cells were examined by overexpressing and RNAi technology.The results showed that the expression of Rab7 had no eff ect on NDV infection while overexpression of Rab5 and Rab11 promoted NDV infection,indicating that Rab5and Rab11 rather than Rab7 regulated NDV transport.This study enriched the knowledge on the pathways and intracellular transport of NDV invasion into host immune cells and provided a theoretical basis for comprehensively elucidating the mechanism of NDV infection.

关 键 词：新城疫病毒 HD11细胞 小窝蛋白介导的内吞 RAB蛋白 

分 类 号：S858.31[农业科学—临床兽医学]