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汉滩病毒(HTNV)感染正常人肺上皮细胞及其固有免疫与代谢改变

Infection of human normal lung epithelial cells by Hantaan virus(HTNV)and its impact on innate immunity and metabolism
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摘要 目的确定汉滩病毒(HTNV)可感染BEAS-2B正常人肺上皮细胞并通过转录组分析HTNV感染诱导的宿主免疫应答和代谢改变。方法采用蛋白免疫印迹、实时荧光定量PCR和免疫荧光实验检测BEAS-2B细胞内的病毒载量,并使用RNA测序技术进行转录组分析。结果在HTNV感染的BEAS-2B细胞中,HTNV核衣壳蛋白(NP)和小片段(S)表达均随时间增加。对HTNV感染BEAS-2B细胞48 h的转录组数据分析,得到328个差异基因,基因本体论(GO)富集及京东基因与基因组百科全书(KEGG)富集分析发现其差异主要集中在干扰素应答及固有免疫模式识别受体相关通路。通过蛋白质互作网络分析发现多个固有免疫应答相关基因,此外筛选到4个编码去整合素和具有血小板反应蛋白基序的金属蛋白酶的基因。通过对代谢通路分析发现3个与萜类骨架生物合成相关的基因,2个与糖酵解/糖异生相关基因和2个类固醇激素生物合成相关基因。此外,差异基因的亚细胞定位分析表明多个差异基因位于线粒体。结论HTNV能有效感染BEAS-2B细胞,可作为体外HTNV感染人肺上皮的细胞模型。生物信息学方法筛选的HTNV感染相关的差异基因及代谢通路,可为研究HTNV感染的分子机制提供理论依据。 Objective To confirm that Hantaan virus(HTNV)can infect BEAS-2B human normal lung epithelial cells and examine the host immune response and metabolic changes induced by HTNV infection by transcriptomic analysis.Methods Western blotting,quantitative real-time PCR and immunofluorescence assay were used to assess the viral load in BEAS-2B cells,and RNA sequencing was employed for transcriptomic analysis.Results Following the infection of BEAS-2B cells with HTNV,there was an increase in the expression of HTNV nucleocapsid protein(NP)and small segment(S)over time.A transcriptomic analysis of these infected cells at 48-hour mark identified 328 genes that were differentially expressed.GO and KEGG enrichment analysis revealed that these differences were primarily associated with interferon response and innate immune pattern recognition receptor pathways.Protein-protein interaction network analysis identified several genes related to innate immune responses,including four genes encoding disintegrin and metalloproteinase with thrombospondin motifs.Metabolic pathway analysis showed three genes related to terpenoid backbone biosynthesis,two genes related to glycolysis/gluconeogenesis and two genes related to steroid hormone biosynthesis.Subcellular localization analysis indicated that many of the differentially expressed genes were located in mitochondria.Conclusion HTNV is capable of effectively infecting BEAS-2B cells,making them a suitable in vitro model for studying HTNV infection in human lung epithelial.By utilizing bioinformatics methods to screen for differentially expressed genes and metabolic pathways associated with HTNV infection,researchers can establish a theoretical foundation for investigating the molecular mechanisms underling HTNV infection.
作者 丁亚鑫 侯诗源 孙丹妮 康华瑞 马晓晗 刘梓谕 刘蓉蓉 吴兴安 DING Yaxin;HOU Shiyuan;SUN Danni;KANG Huarui;MA Xiaohan;LIU Ziyu;LIU Rongrong;WU Xingan(College of Life Science,Northwest University,Xi’an 710069;Department of Microbiology and Pathogenic Biology,Basic Medical Science Academy,Air Force Medical University,Xi’an 710032,China)
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2024年第5期385-394,共10页 Chinese Journal of Cellular and Molecular Immunology
基金 国家自然科学基金(81971563) 陕西省自然科学基础研究计划重点项目(2024JC-ZDXM-42)。
关键词 汉滩病毒(HTNV) 生物信息学 固有免疫应答 代谢 Hantaan virus(HTNV) bioinformatics innate immune response metabolism
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