摘要
为了研究猪瘟活疫苗(兔源)免疫猪体后的免疫应答,本试验选择未免疫猪瘟疫苗且猪瘟抗原抗体阴性的4周龄小猪10头,随机分成2个组别,分别免疫猪瘟活疫苗(兔源)、健康家兔脾淋组织(组织对照),采集免疫后3 d的前腔静脉血液样本,利用RNA-Seq技术进行转录组测序分析,并采用荧光定量PCR(RT-qPCR)方法验证。结果表明:猪瘟活疫苗(兔源)免疫组与脾淋组织对照组相比,外周血中差异表达基因有317个(P<0.05),其中上调表达的基因数量为221个,下调表达的有96个;差异基因GO功能富集分析显示,与生物过程相关的有483个,细胞组分相关的98个,分子功能相关的有87个,其中显著差异转录的mRNA主要参与对病毒的防御反应(defense response to virus)、病毒过程(viral process)等生物过程;KEGG信号通路富集分析表明,涉及的通路数量有215个,与免疫和抗炎相关的有甲型流感(Influenza A)、EB病毒感染(Epstein-Barr virus infection)、RIG-I样受体(RIG-I-like receptor signaling pathway)、TNF信号通路(TNF signaling pathway)、NOD-like受体信号通路(NOD-like receptor signaling pathway)等,其中CXCL10、RSAD2、OAS1、OAS2、MX1、ISG15等基因上调表达;随机筛选了CXCL10、NOS3、CD86、NOD14个差异表达基因进行RT-qPCR验证,结果显示RT-qPCR和测序结果间的差异表达基因趋势基本一致,说明猪瘟活疫苗(兔源)免疫3 d可刺激猪体先天性免疫应答。
In order to investigate the immunity response of pigs immunized live classical swine fever vaccine(rabbit origin),ten fourweek-old piglets were divided into 2 groups randomly and immunized with live classical swine fever vaccine(rabbit origin)and rabbit spleen and lymph tissue(control group),respectively.The peripheral blood samples were collected on the 3rd day post immunization.The transcriptome profiles of blood samples were examined by RNA-seq technology and verified by quantitative fluorescent PCR(RT-qPCR).The results showed that there were 317 differentially expressed genes(P<0.05)detected in vaccinated piglets as compared with the control group,among which 221 genes were up-regulated and 96 genes were down-regulated.GO function enrichment analysis of differential genes showed 483 genes related to biological processes,98 genes to cellular components,and 87 genes to molecular functions.Among them,the significantly and differentially transcribed mRNAs were mainly involved in biological processes such as defense response to virus and viral process.KEGG signaling pathway enrichment analysis showed 215 pathways involved.Several pathways were related to immunity and anti-inflammation effect such as influenza A,Epstein-Barr virus infection,RIG-I-like receptor,TNF and nod-like receptor signaling pathways etc.Among them,CXCL10,RSAD2,OAS1,OAS2,MX1,ISG15 genes were up-regulated and CXCL10,NOS3,CD86 and NOD1 were randomly screened by RT-qPCR.These results showed that the trend of differentially expressed genes was basically consistent between RT-qPCR and sequencing,suggesting that the live classical swine fever vaccine(rabbit origin)stimulated the innate immune response of piglets.
作者
包松英
庄许诺
江兴华
王全溪
BAO Songying;ZHUANG Xunuo;JIANG Xinghua;WANG Quanxi(Jofunhwa Biotechnology(Fuzhou)Co.,Ltd.,Fuzhou 350014,China;Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health,Fujian Agriculture And Forestry University,Fuzhou 350002,China;University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province,Fuzhou 350002,China)
出处
《中国动物传染病学报》
CAS
北大核心
2022年第2期48-55,共8页
Chinese Journal of Animal Infectious Diseases
基金
福建省重大专项专题项目(2019NZ09007)。
