摘要
伪狂犬病病毒(PRV)的天然宿主是猪,但是能够感染多种哺乳动物。PRV是一种嗜神经性α疱疹病毒,小鼠感染PRV后会产生严重的神经症状和神经炎症。为了探究小鼠感染PRV后其背根神经节(DRG)的天然免疫应答情况,本研究将PRV TJ株以10^(6)pfu剂量接种小鼠,在濒死期剖杀小鼠,取其DRG进行转录组测序,并使用DESeq2软件对测序结果分析,结果显示,与对照组相比,PRV感染小鼠的DRG内共有6 502条基因的转录水平发生改变,其中转录水平升高的基因有3 703条,降低的基因有2 799条(差异倍数>2,且P≤0.05)。利用clusterProfiler R软件对发生差异转录的基因进行了KEGG分析,结果显示,转录水平发生差异的基因参与细胞内20种信号通路途径,其中12条信号通路与天然免疫反应有关,提示PRV感染小鼠后会激活DRG内广泛的天然免疫反应。差异转录基因数量最多的5条天然免疫信号通路分别是丝裂原活化蛋白激酶(MAPK)信号通路、核苷酸结合寡聚化结构域样受体(NLR)信号通路、肿瘤坏死因子(TNF)信号通路、细胞凋亡(Apoptosis)信号通路和信号转导及转录激活因子(JAK-STAT)信号通路,上述天然免疫反应通路可能在PRV诱导的神经炎症方面有重要作用。本研究随机选择了10条转录水平存在差异的基因,采用荧光定量PCR验证,结果显示其变化趋势均与转录组测序数据一致。本研究首次分析了PRV感染小鼠DRG内转录组的变化,为研究PRV感染后诱导宿主的天然免疫反应相关信号通路提供了参考,并为PRV致病机制等相关研究奠定了基础。
Pseudorabies virus(PRV),a virus whose natural host is the pig,can infect a wide range of mammals.PRV is a neurotropic alphaherpesvirus.The mice infected with PRV develop severe neurological symptoms leading to neuroinflammation.To explore the immune response in peripheral nervous system(PNS)of mice,the mice were infected with 10^(6) pfu PRV TJ strain and euthanized at the moribund state.Dorsal root ganglia(DRG)were collected,and the transcriptomes were sequenced.The data was analyzed by using DESeq2 software.The results showed that a total of 6502 genes were differentially transcribed in the experimental group compared with the blank control group,among them 3703 genes were upregulated and 2799 genes were downregulated(fold changes>2,and P≤0.05).Systematic bioinformatics analysis with Kyoto Encyclopedia of Genes and Genomes(KEGG)by using cluster Profiler R software revealed that differentially transcribed genes were involved in 20 signaling pathways,and 12 signaling pathways of those were associated with innate immunity responses,which maybe induce broad innate immune response after PRV infection.Mitogen-activated protein kinase(MAPK)signaling pathway,nucleotide-binding oligomerization domain-like receptor(NLR)signaling pathway,tumor necrosis factor(TNF)signaling pathway,apoptosis signaling pathway and signal transducers and activators of transcription(JAK-STAT)signaling pathway were the top 5 innate immune signaling pathways that showed the largest number of differentially transcribed genes.These pathways have an important role in antiviral innate immune response.Besides,10 differentially transcribed genes were randomly selected for verification by real-time PCR,and the changing trend was consistent with that of the transcriptome data.Taken together,the transcriptomic data set is able to provide a reference for the studies of pathogenicity and innate immune response induced by PRV in DRGs,and provide important basic data and research direction for further understanding the pathogenic mechanisms of PRV infectio
作者
王冰
吴红霞
李淼
高莹
袁梦淇
仇华吉
孙元
WANG Bing;WU Hong-xia;LI Miao;GAO Ying;YUAN Meng-qi;QIU Hua-Ji;SUN Yuan(State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2022年第6期591-597,共7页
Chinese Journal of Preventive Veterinary Medicine
基金
国家自然科学基金面上项目(31870152)
黑龙江省自然科学基金面上项目(C2018067)。
关键词
伪狂犬病病毒
小鼠
背根神经节
转录组测序
差异转录基因
pseudorabies virus
mice
dorsal root ganglia
transcriptome sequencing
differentially transcribed genes
