ALV-J下调鸡MARCO转录水平拮抗宿主抗病毒天然免疫应答的研究  

ALV-J antagonizes host antiviral innate immune response by downregulating the transcription levels of chicken macrophage receptor MARCO

作　　者：陈绪靖 吉紫荆 赵宗仪 豆春峰 柴文娴 陈世豪[1] 耿拓宇[1] 胡序明[1] 崔恒宓[1] CHEN Xujing;JI Zijing;ZHAO Zongyi;DOU Chunfeng;CHAI Wenxian;CHEN Shihao;GENG Tuoyu;HU Xuming;CUI Hengmi(Institute of Epigenetics and Epigenomics/Joint International Research Laboratory of Agricultural&Agri-Product Safety of the Ministry of Education of China/College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China;Changzhou Animal Disease Prevention and Control Center,Changzhou 213002,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/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 [2]常州市动物疫病预防控制中心,江苏常州213002

出　　处：《扬州大学学报（农业与生命科学版）》2021年第2期46-51,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金重大研究项目(91540117);国家自然科学基金资助项目(81773013);常州市科技局重点科研项目(CE20202033);江苏省优势学科建设工程项目(PAPD);江苏省高校大学生创新创业训练计划项目(x20180581)。

摘　　要：为研究鸡巨噬细胞受体(MARCO)基因与J亚群禽白血病病毒(ALV-J)感染两者间的关联,先过表达ALV-J env基因不同结构域后通过RT-qPCR检测MARCO基因mRNA的转录水平,再构建MARCO真核表达载体并转染HD11细胞,通过RT-qPCR和Western blot检测MARCO过表达对ALV-J复制水平的影响,最后通过RT-qPCR检测MARCO过表达对巨噬细胞天然免疫相关基因TLR3和IFIH1转录水平的影响。结果表明:ALV-J env蛋白对MARCO基因mRNA转录水平有极显著抑制作用;过表达MARCO极显著抑制ALV-J病毒的复制;过表达MARCO后,TLR3和IFIH1基因mRNA转录水平均极显著上调。这一研究揭示了MARCO基因在ALV-J感染巨噬细胞后的动态转录水平规律及其抑制ALV-J增殖的功能,其可能通过诱导天然免疫基因TLR3、IFIH1高转录水平的途径来实现抗病毒作用,为今后研究MARCO的功能及治疗禽白血病提供了新的思路。To explore the association between chicken macrophage receptor with collagenous structure(MARCO) gene and subgroup J avian leukemia virus(ALV-J) infection. The transcription level of MARCO gene mRNA was detected by RT-qPCR after overexpressing the different domains of ALV-J env gene in the present study, then the MARCO eukaryotic expression vector was constructed and transfected into HD11 cells, the effect of MARCO overexpression was tested on the replication level of ALV-J by RT-qPCR and Western blot, finally, RT-qPCR was performed to detect the effect of MARCO overexpression on the transcriptional levels of TLR3 and IFIH1 that are the genes related to innate immunity of macrophages. The results showed that ALV-J env protein had a very significant inhibitory effect on the MARCO gene mRNA transcription level(P<0.01);ALV-J replication could be obviously restrained by over-expressing of MARCO(P<0.01);TLR3 and IFIH1 gene mRNA transcription levels were significantly up-regulated after overexpression of MARCO(P<0.01).This study revealed the dynamic transcription level of MARCO gene in macrophages after ALV-J infection and its effect on inhibiting the proliferation of ALV-J,and it may be achieved by inducing high transcription levels of innate immune genes TLR3 and IFIH1,which laid a foundation for the study of the function of MARCOand the treatment of avian leukemia in the future.

关 键 词：MARCO基因 巨噬细胞 天然免疫 基因克隆 J亚群禽白血病病毒 

分 类 号：S831.2[农业科学—畜牧学] Q78[农业科学—畜牧兽医]