畜牧与兽医2024，Vol.56Issue(8)：59-69,11.
伪狂犬病病毒感染猪肺泡巨噬细胞和小肠上皮细胞的转录组动态分析
Transcriptome dynamics of pseudorabies virus-infected porcine alveolar macrophages and small intestinal epithelial cells
摘要
Abstract
To investigate the interaction of pseudorabies virus(PRV)with porcine alveolar macrophages and small intestinal epithelial cells,PRV was infected with porcine alveolar macrophage cell line(3D4/21)and small intestinal epithelial cell line(IPEC-J2);the viral titer and viral gE gene copy number were measured at each time point from 0-48 h after the infection;the cytopathic lesions(CPE)and im-munofluorescence were observed,and RT-qPCR,ELISA for tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6).Transcriptome sequencing(RNA-seq)analyses were performed on the samples at 0,6,12 and 18 h after the PRV infection.The results showed that,when PRV infected 3D4/21 cells and IPEC-J2 cells for 96 h,the viral titer was 10-8.16 TCID50/0.1 mL and 10-7.76 TCID50/0.1 mL,respectively;the viral gE gene copy number reached the maximum at 18 and 24 h,respectively;the CPE began to appear at 10 and 12 h,respectively;the viral protein expression increased gradually with the infection.The expression of the viral proteins gradually increased with the prolongation of infection time;the expression of TNF-α,IL-6 and IL-1β gradually increased after the PRV in-fection;3D4/21 cells and IPEC-J2 cells were analyzed by RNA-seq at four time points,namely,0,6,12 and 18 h of the PRV infection,and the differentially expressed genes were screened out at 8 313,6 999,6 693,5 714,among which the significantly altered genes of Ras off structural domain family 6(RASSF6),zinc finger protein 667(ZNF667),kinin superfamily protein 3C(KIF3C),and early-response gene 3(IER3)might be related to cellular inflammatory response,and the RT-qPCR validation was in agreement with the RNA-seq re-sults.The gene ontology(GO)analysis showed that the differential genes were enriched in cellular processes,bioregulation,and regulation of cellular metabolism,etc.the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment showed that the differential genes were mainly involved in the cancer pathway,the mitogen-activated protein kinase(MAPK)signaling pathway,and Toll-like receptors(TLRs)signaling pathway,etc.,among which,the significantly altered genes of the Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(MYD88)-nuclear transcription factor кB(NF-кB)pathway involved in inflammation were in general agreement with the trend of the KEGG pathway analysis data.To conclude,PRV infection affected the inflammatory response of 3D4/21 cells and IPEC-J2 cells,and this study laid the foundation for further exploration of the pathogenic mechanism of PRV.关键词
3D4/21细胞/IPEC-J2细胞/伪狂犬病病毒/差异表达基因Key words
3D4/21 cell/IPEC-J2 cell/pseudorabies virus/differentially expressed genes分类
农业科技引用本文复制引用
张王芝,舒相华,张莹,黄鑫,储宗秀,宋春莲..伪狂犬病病毒感染猪肺泡巨噬细胞和小肠上皮细胞的转录组动态分析[J].畜牧与兽医,2024,56(8):59-69,11.基金项目
云南省农业重大科技专项(202202AE090027) （202202AE090027）
云南省乡村振兴科技专项(202304BI090011) （202304BI090011）
