鸡巨噬细胞CCL4抗禽白血病病毒作用及其机制初探  

A preliminary study on the effect and mechanism of CCL4against avian leukemia virus in chicken macrophage cell(HD11)

作　　者：陈绪靖 赵宗仪 吉紫荆 豆春峰 吴挺 陈世豪[1,2] 耿拓宇 胡序明[1,2] 崔恒宓 CHEN Xujing;ZHAO Zongyi;JI Zijing;DOU Chunfeng;WU Ting;CHEN Shihao;GENG Tuoyu;HU Xuming;CUI Hengmi(Institute of Epigenetics and Epigenomics,College of Animal Science and Technology,Yangzhou University,Yangzhou,Jiangsu 225009,China;Joint International Research Laboratory of Agricultural&Agri-Product Safety of the Ministry of Education of China,Yangzhou University,Yangzhou,Jiangsu 225009,China)

机构地区：[1]扬州大学动物科学与技术学院/表观遗传学及表观基因组学研究所,江苏扬州225009 [2]扬州大学教育部农业与农产品安全国际合作联合实验室,江苏扬州225009

出　　处：《中国兽医学报》2021年第4期670-675,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金面上资助项目(91540117,81773013);江苏省畜牧学优势学科资助项目;江苏省高等学校大学生创新创业训练计划资助项目(x20180581)。

摘　　要：为了探究CCL4基因抗J亚群禽白血病病毒(ALV-J)的作用机制,观察了CCL4基因在ALV-J感染鸡巨噬细胞(HD11)不同时间(6,12,24,36,48h)后的表达规律,分别构建CCL4、ALV-J env、ALV-J GP85和ALV-J GP37真核表达载体并转染HD11细胞,通过RT-qPCR检测CCL4过表达对ALV-J复制水平及CCL4受体CCR5和其下游天然免疫相关基因及其他抗病毒基因的表达水平的影响,同时分析ALV-J env对CCL4基因表达的影响。结果显示:ALV-J感染HD11细胞后主要抑制CCL4表达(P<0.01);ALV-J env蛋白对CCL4有显著抑制作用(P<0.01);而过表达CCL4显著抑制了ALV-J病毒复制(P<0.01);CCL4可能通过激活抗病毒基因S100A9及其上、下游通路抑制ALV-J病毒增殖。本研究揭示了CCL4在病毒感染后巨噬细胞中的动态表达规律及其抗ALV-J作用,为今后研究CCL4的功能及其宿主遗传抗性奠定基础。The purpose of this study is to explore the essential role of CCL4in the process of anti-J subgroup avian leukosis virus(ALV-J).The expression level of the CCL4gene was observed in chicken macrophage cell(HD11)infected with ALV-J at different times(6,12,24,36and 48h).CCL4,ALV-J env,ALV-J GP85and GP37eukaryotic expression vectors were constructed and transfected into HD11cells.RT-qPCR was performed to detect the expression of these genes and the influence of overexpression of CCL4on ALV-J replication level and expression changes of CCL4receptor gene CCR5,immune-related genes and other antiviral genes.Then,the influence of ALV-J env expression on CCL4was further analyzed.The results showed that ALV-J infection significantly repressed the expression of CCL4(P<0.01);ALV-J env protein had a significant inhibitory effect on CCL4expression(P<0.01).However,over-expressing of CCL4could restrain ALVJ replication(P<0.01).CCL4may control ALV-J proliferation by activating antiviral gene S100A9 and its upstream or downstream signal pathways.This study reveals the dynamic expression of CCL4in macrophage cells infected with ALV-J and its anti ALV-J effect,which lays a foundation for further study of the function of CCL4and host genetic resistance.

关 键 词：CCL4 巨噬细胞 天然免疫 ALV-J 

分 类 号：S852.65[农业科学—基础兽医学]