鸡胚免疫新城疫疫苗后宿主法氏囊转录组学分析  

Transcriptomic Analysis of Host’s Bursa of Fabricius After in-ovo Vaccination with Newcastle Disease Vaccine

作　　者：蒋立人 冯贺龙 姜鑫瑞 王梓晨 张高峰 商雨[2] 曾哲[2] 姚伦 汪宏才[2] 罗青平[2,3] 田光明 温国元[2] JIANG Liren;FENG Helong;JIANG Xinrui;WANG Zichen;ZHANG Gaofeng;SHANG Yu;ZENG Zhe;YAO Lun;WANG Hongcai;LUO Qingping;TIAN Guangming;WEN Guoyuan(College of Animal Science and Technology,Yangtze University,Jingzhou 434025,China;Key Laboratory of Prevention and Control Agents for Animal Bacteriosis(Ministry of Agriculture and Rural Affairs),Hubei Provincial Key Laboratory of Animal Pathogenic Microbiology,Institute of Animal Sciences and Veterinary Medicine,Hubei Academy of Agricultural Sciences,Wuhan 430064,China;Hubei Hongshan Laboratory,Wuhan 430070,China)

机构地区：[1]长江大学动物科学技术学院,荆州434025 [2]湖北省农业科学院畜牧兽医研究所,农业农村部畜禽细菌病防治制剂创制重点实验室&畜禽病原微生物学湖北重点实验室,武汉430064 [3]湖北洪山实验室,武汉430070

出　　处：《中国畜牧兽医》2024年第4期1632-1641,共10页China Animal Husbandry & Veterinary Medicine

基　　金：湖北省重点研发计划项目(2023BBB034);国家肉鸡产业技术体系(CARS-41);湖北省农业科技创新中心项目(2019-620-000-001-17)。

摘　　要：【目的】通过转录组学分析鸡胚免疫新城疫病毒(Newcastle disease virus,NDV)TS09-C株后宿主法氏囊内免疫关键基因及信号通路分子转录差异,探究免疫后宿主法氏囊内免疫相关分子的调控反应。【方法】将60只18胚龄SPF鸡胚随机分为免疫组和对照组,30枚/组,免疫组将NDV TS09-C株通过羊膜腔接种鸡胚,剂量为103.0 EID 50/枚,注射体积为0.1 mL,对照组以同样的方法接种等体积PBS。检测免疫后不同时间点肺脏、胸腺、脾脏和法氏囊组织的病毒载量。选取病毒载量最高的法氏囊组织进行转录组学测序后,筛选免疫相关差异表达基因并分析其参与的主要功能及通路。【结果】免疫后1 d法氏囊组织中即可监测到病毒,免疫后3 d病毒载量达到最高。将免疫后3 d的法氏囊进行转录组学分析,与对照组相比,免疫组有453个基因上调、98个基因下调,其中免疫相关基因有36个上调、10个下调。免疫相关差异表达基因主要富集于自噬和细胞因子-细胞因子受体相互作用的免疫相关通路。差异表达基因蛋白互作分析显示,自噬通路中富集的BCL2与其他免疫相关基因有较多互作关系,且在B细胞活化、分化、免疫系统发育等功能注释中均有参与。【结论】本研究结果揭示了鸡胚免疫NDV TS09-C株后法氏囊内调控B细胞活化、分化的潜在靶向基因及通路,为进一步研究该毒株鸡胚免疫后B淋巴细胞调控的分子机制提供新的信息。【Objective】The aim of this experiment was to analyze the transcriptional differences of immune key genes and signaling pathway molecules in host’s bursa of Fabricius after in-ovo immunization with Newcastle disease virus TS09-C strain in chicken embryos using transcriptomics,and explore the immune related molecular regulatory responses in the host’s bursa of Fabricius after immunization.【Method】Sixty 18 embryo SPF chicken embryos were randomly divided into immunization group and control group,with 30 embryos per group.The immunization group inoculated NDV TS09-C strain into chicken embryos through the amniotic cavity,the dose was 103.0 EID 50/piece and the injection volume was 0.1 mL,while the control group was inoculated with equal volume PBS using the same method.The viral load of lung,thymus,spleen and bursa of Fabricius tissue was detected at different time points after immunization.The bursa of Fabricius tissue with the highest viral load was selected for transcriptome sequencing and analysis.Immune related differentially expressed genes were screened and the main functions and pathways they involved were analyzed.【Result】The virus could be detected in bursa of Fabricius tissue 1 day after immunization,and the viral load reached the highest level 3 days after immunization.Transcriptome analysis was performed on the bursa of Fabricius 3 days after immunization.Compared with control group,there were 453 up-regulated genes and 98 down-regulated genes in immunization group,including 36 up-regulated genes and 10 down-regulated genes related to immunity.Immune related differentially expressed genes were mainly enriched in the immune related pathways of autophagy and cytokine-cytokine receptor interactions.Differentially expressed gene protein-protein interaction analysis showed that BCL2 enriched in the autophagy pathway interacted with other immune related genes,and was involved in functional annotation such as B cell activation,differentiation,and immune system development.【Conclusion】The res

关 键 词：新城疫病毒(NDV) TS09-C株 鸡胚免疫 法氏囊 转录组分析 

分 类 号：S852.65[农业科学—基础兽医学]