新城疫疫苗免疫武定鸡和霞烟鸡的外周血淋巴细胞转录组分析  

Transcriptome Analysis of Peripheral Blood Lymphocytes of Wuding Chicken and Xiayan Chicken Immunized with Newcastle Disease Vaccine StrainⅣ

作　　者：陈曦 刘通 白潜 叶玮琪 和凤平 向斌 张立梅 江康峰 李小兵 杨亮宇 CHEN Xi;LIU Tong;BAI Qian;YE Weiqi;HE Fengping;XIANG Bin;ZHANG Limei;JIANG Kangfeng;LI Xiaobing;YANG Liangyu(College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201)

机构地区：[1]云南农业大学动物医学院,云南昆明650201

出　　处：《中国家禽》2022年第7期43-49,共7页China Poultry

基　　金：云南省万人计划-产业技术领军人才(A3012020032);国家自然科学基金地区科学基金项目(32060818);现代农业产业技术体系建设专项资金(CARS-40-S25)。

摘　　要：为了探究武定鸡和霞烟鸡免疫新城疫疫苗后外周血淋巴细胞转录组的变化,试验随机选取60日龄健康武定鸡和霞烟鸡各5只,在免疫新城疫Ⅳ系疫苗前和免疫后第14天采血并分离其外周血淋巴细胞。提取外周血淋巴细胞总RNA,进行mRNA转录组测序,将测序获得的数据基于FPKM、GO分析、KEGG数据库功能注释,比较免疫前后外周血淋巴细胞差异表达基因和信号通路;RT-qPCR验证随机挑选的差异基因的相对表达量。结果显示:在武定鸡和霞烟鸡外周血淋巴细胞中分别获得5409万条和5095万条高质量序列;武定鸡免疫前后外周血淋巴细胞转录组差异基因共524个,其中上调基因296个、下调基因228个;霞烟鸡免疫前后差异基因178个,其中上调135个、下调43个;武定鸡差异基因显著富集于炎症反应、IL-6以及趋化因子介导的信号通路的生物过程,霞烟鸡则显著富集于基底胶原蛋白调节、细胞膜成分、血管生成等的生物过程;武定鸡和霞烟鸡免疫前后差异基因均富集于IL-17信号通路和类风湿性关节炎信号通路;武定鸡与霞烟鸡在接种疫苗后IL-8与TNF-α表达量均显著上调,武定鸡表达量显著高于霞烟鸡。研究表明接种新城疫Ⅳ系疫苗后,武定鸡与霞烟鸡免疫前后差异表达基因分别富集于炎症通路和血管生成通路,新城疫疫苗对武定鸡和霞烟鸡的外周血淋巴细胞调节途径存在差异。In order to investigate the changes of lymphocyte transcriptome in peripheral blood of Wuding chicken and Xiayan chicken immunized with Newcastle disease vaccine,5 healthy Wuding chickens and 5 Xiayan chickens aged 60 days were randomly selected and their peripheral blood lymphocytes were collected before and 14 days after immunization with Newcastle disease vaccine strainⅣ.Total RNA was extracted from peripheral blood lymphocytes for mRNA transcriptome sequencing.The data obtained by sequencing were compared based on FPKM,GO analysis and KEGG database functional annotation to compare the differentially expressed genes and signaling pathways in peripheral blood lymphocytes before and after immunization.RT-qPCR verified the relative expression levels of randomly selected differential genes,and the results showed that 54.09 million and 50.95 million clean reads were obtained respectively from peripheral blood lymphocytes of Wuding chicken and Xiayan chicken.There were 524 differentially expressed genes(296 up-regulated genes and 228 down-regulated genes)in peripheral blood lymphocytes transcriptome of Wuding chicken before and after immunization.One hundred seventy eight differential genes were up-regulated 135 and down-regulated 43 before and after immunization in Xiayan chicken.The differential genes of Wuding chicken were significantly enriched in the biological process of inflammatory response,IL-6 and chemokine mediated signaling pathway,while those of Xiayan chicken were significantly enriched in the biological process of angiogenesis.The differential genes were enriched in IL-17 signaling pathway and rheumatoid arthritis signaling pathway in Wuding and Xiayan chickens before and after immunization.The expression levels of IL-8 and TNF-αwere significantly up-regulated in Wuding chicken and Xiayan chicken after vaccination,and the expression levels of IL-8 and TNF-αin Wuding chicken were significantly higher than that in Xiayan chicken.The results showed that the differentially expressed genes of Wuding ch

关 键 词：武定鸡 霞烟鸡 新城疫疫苗 转录组测序 

分 类 号：S852.65[农业科学—基础兽医学]