长非编码RNA lnc-ALVE1-AS1与禽白血病病毒抗性和易感品系鸡相关性分析  

Correlation Between Long Non-Coding RNA lnc-ALVE1-AS1 and Avian Leukemia Virus Resistant and Susceptible Chicken Strains

作　　者：柴文娴 骆欢 陈绪靖 陈世豪[2,3] 耿拓宇 崔恒宓[2,3] 胡序明 CHAI Wenxian;LUO Huan;CHEN Xujing;CHEN Shihao;GENG Tuoyu;CUI Hengmi;HU Xuming(Changzhou Animal Disease Prevention and Control Center,Changzhou 213002,China;Institute of Epigenetics and Epigenomics,College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China;Joint International Research Laboratory of Agricultural&Agri-Product Safety,Ministry of Education of China,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]常州市动物疫病预防控制中心,常州213002 [2]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,扬州225009 [3]扬州大学教育部农业与农产品安全国际合作联合实验室,扬州225009

出　　处：《中国动物传染病学报》2023年第4期26-32,共7页Chinese Journal of Animal Infectious Diseases

基　　金：常州市科学技术局重点项目(CE20202033);国家自然科学基金(31602032);国家自然科学基金重大研究计划(91540117);江苏省畜牧学优势学科项目。

摘　　要：内源性反转录病毒是长非编码RNA的重要来源,在抗病毒先天性免疫中发挥重要作用。鸡内源性反转录病毒衍生的长非编码RNA lnc-ALVE1-AS1能够激活细胞免疫功能并抑制禽白血病病毒增殖。为进一步探讨lnc-ALVE1-AS1在ALV抗性和易感品系鸡中的表达规律及其与禽白血病病毒抗性的相关性,本研究通过荧光定量PCR发现,lnc-ALVE1-AS1在ALV抗性(G1)品系鸡免疫器官(胸腺、脾脏和法氏囊)和CEF细胞中的表达水平显著高于ALV易感(G3)品系鸡。病毒感染实验表明lnc-ALVE1-AS1在ALVJ感染CEF(G1和G3来源)细胞24h和96h后均显著下调;但是,lnc-ALVE1-AS1在G3来源CEF细胞中的表达水平下降更为明显,且ALVJ病毒复制水平显著高于G1鸡来源的CEF细胞。进一步通过lnc-ALVE1-AS1过表达实验证实,lnc-ALVE1-AS1不仅可以显著上调G3来源CEF细胞dsRNA识别受体(TLR3和IFIH1)和抗病毒天然免疫基因(IFI27-L2、IFIT5、IFITM3、IFN-β、ISG12-2和RASD2)表达,还显著抑制ALVJ复制。本研究揭示了lnc-ALVE1-AS1在禽白血病病毒抗性和易感品系鸡细胞和免疫组织器官中的表达规律和抗病毒功能,为抗病育种研究提供了新的遗传基础。Previous studies have shown that endogenous retroviruses are one of the important sources of long non-coding RNA and play an important role in antiviral innate immunity.It has been reported that the long non-coding RNA lnc-ALVE1-AS1 derived from chicken endogenous retrovirus can activate cellular immune function and inhibit the proliferation of Avian leukemia virus.In order to further understand and explore the expression pattern of lnc-ALVE1-AS1 in ALV-resistant and susceptible chicken strains and its correlation with host avian leukemia virus resistance,this study revealed that the expression level of lnc-ALVE-AS1 in ALV resistant(G1) chicken immune organs(thymus,spleen and bursa of fabricius) and chicken embryo fibroblast CEF was significantly higher than that of susceptible(G3) chicken as detected in the strand-specific fluorescence quantitative PCR.The results of viral infection experiments showed that the expression of lnc-ALVE1-AS1 was significantly down-regulated in ALV resistant G1 and susceptible G3 chicken CEF cells infected with ALVJ virus after 24 h and 96 h.However,the decline level of lnc-ALVE1-AS1 expression was more obvious in ALV-susceptible G3 chicken CEF cells while the ALVJ virus replication level was significantly higher than that of ALV-resistant G1 chicken CEF cells.The overexpression experiments further confirmed that lnc-ALVE1-AS1 significantly upregulated the expression levels of dsRNA recognition receptors(TLR3 and IFIH1) and antiviral innate immune genes(IFI27-L2,IFIT5,IFITM3,IFN-β,ISG12-2 and RASD2) and inhibited ALVJ replication in ALV-susceptible G3 chicken CEF cell.In summary,this study represented the expression pattern of lnc-ALVE1-AS1 in the ALV-resistant and susceptible chicken cells and immune organs and its antiviral function,which provided a new genetic basis for disease resistance breeding research.

关 键 词：内源性反转录病毒 lnc-ALVE1-AS1 禽白血病病毒 鸡胚成纤维细胞 

分 类 号：S858.31[农业科学—临床兽医学]