BCG感染牛肺泡巨噬细胞的转录组测序和分析

Transcriptome sequencing and analysis of bovine alveolar macrophages infected with BCG

目的:通过卡介苗(BCG)感染牛肺泡巨噬细胞(BAM)的转录组测序及生物信息学分析,揭示差异表达的基因和长链非编码RNA (lncRNA),为巨噬细胞抗结核分枝杆菌感染的免疫调控机制研究提供理论依据。方法:采用肺脏灌洗离心法收集并培养BAM,分为感染组和未感染组,感染组经BCG感染后12h,利用转录组测序技术(RNA-Seq)检测感染组和未感染组BAM mRNA表达谱及lncRNA表达谱,并进行生物信息学相关分析。结果:与未感染组比较,感染组BAM差异表达的lncRNAs共有119个(P<0.05),差异表达的mRNA共有1 111个(P<0.05)。在差异表达基因的功能富集分析中,最显著富集项与免疫功能相关(GO:0006955,P<0.05),共有125个基因,其中有63个基因表达上调,62个基因表达下调,且白细胞介素6(IL-6)、白细胞介素17 (IL-17)和白细胞介素23A (IL-23A)等促炎因子表达上调。lncRNA靶基因预测,差异表达的lncRNA参与转化生长因子β(TGF-β)信号通路及ABC转运体信号通路的调控(P<0.05)。结论:BCG感染BAM后,激发宿主细胞产生强烈的免疫反应,引起lncRNA和mRNA表达谱的改变。

Objective:To reveal the differentially expressed genes and long non-coding RNAs(lncRNAs)by sequencing the transcriptome of bovine alveolar macrophages(BAM)infected with Bacillus Calmette-Guérin(BCG)and analyzing their bioinformations,and to provide theoretical foundation for the research on immune regulation mechanism of anti-infection of Mycobacterium tuberculosis of macrophages.Methods:The BAM were collected by pulmonary lavage and centrifugation and cultured and divided into infected group and uninfected group.After infection for 12 h in infected group,the expression profiles of mRNA and lncRNA in infected group and uninfected group were detected by RNA-Seq,and bioinformatics analysis was carried out.Results:compared with uninfected group,there were 119 differentially expressed lncRNAs and 1111 differentially expressed mRNA in infected group(P<0.05).Gene Ontology functional enrichment analysis showed that the most significant enrichment was immune response(GO:0006955,P<0.05),including 125 genes,in which 63 were up-regulated and 62 were down-regulated,and the expressions of proinflammatory factors interleukin-1(IL-6),interleukin-7(IL-7),and interleukin-23A(IL-23A)were up-regulated.Cis target gene prediction and KEGG pathway analysis showed that the differentially expressed lncRNAs were involved in the regulation of transforming growth factor-β(TGF-β)signaling pathway and ATP-binding cassette transporter(ABC transporter)signaling pathway(P<0.05).Conclusion:The host cells are stimulated to produce a strong immune response after the BAM are infected by BCG and results in the changes of lncRNA and mRNA expression profiles.