转录组分析布鲁氏杆菌患者外周血免疫基因的表达情况

Transcriptome analysis of immune genes in the peripheral blood in patients with Brucella infection

目的通过生物信息学分析方法探讨布鲁氏杆菌病的关键失调免疫基因和分子机制。方法从GSE69597中获取布鲁氏杆菌病患者全血转录组表达数据。通过差异分析筛选布鲁氏杆菌病患者和对照之间的差异表达基因(DEGs),并利用immport数据库调取DEGs中免疫相关的基因集(immune-DEGs)。通过Enirchr在线富集工具对immune-DEGs进行富集分析。构建immune-DEGs的蛋白质-蛋白质互作(PPI)网络,并鉴定网络中的高度互联的核心(hub)基因。利用qRT-PCR验证hub基因的表达,并绘制hub基因的ROC曲线。使用ssGSEA算法评估布鲁氏杆菌病患者中免疫细胞的评分,并通过流式细胞术检测血液样本中免疫细胞的水平。结果共获得390个immune-DEGs,富集结果中发现了T细胞受体信号通路和Th17细胞分化等。在10个hub基因中IFNG和TNF在布鲁氏杆菌组中显著高表达。ROC曲线表明IFNG对布鲁氏杆菌病具有良好诊断意义。此外,活化型CD 4 T细胞、效应型CD 4 T细胞、效应记忆型CD 8 T细胞和2型T辅助细胞因子在布鲁氏杆菌患者中明显增多。流式细胞术检测发现与健康对照组相比,布鲁氏杆菌病患者外周血中Th2和Th17细胞比例增高,Th1和Treg细胞比例则降低。结论本研究结果不仅提高了我们对布鲁氏杆菌感染后机体免疫反应的认识,还为诊断和治疗布鲁氏杆菌病提供了更多的方向。

This study aimed to explore the key dysfunctional immune genes and molecular mechanisms of brucellosis by using bioinformatics analysis.Whole blood transcriptome expression data from patients with brucellosis were acquired from GSE69597.Differentially expressed genes(DEGs)between patients with brucellosis and controls were screened through differential analysis,and immune related gene sets(immune-DEGs)in the DEGs were called with the immport database.Enrichment analysis of immune-DEGs was performed with the enirchr online enrichment tool.To construct a protein-protein interaction(PPI)network of immune-DEGs and identify the highly interconnected core(hub)genes in the network,we used qRT-PCR to verify the expression of the hub genes,and we plotted ROC curves of the hub genes.The scoring of immune cells in patients with brucellosis was evaluated with the ssGSEA algorithm,and the levels of immune cells in blood samples were detected by flow cytometry.A total of 390 immune-DEGs were obtained,and the T cell receptor signaling pathway and Th17 cell differentiation were found in the enrichment results.Among the ten hub genes,IFNG and TNF had significantly higher expression in the Brucella group.The ROC curves indicated a good diagnostic value of IFNG for brucellosis.In addition,activated CD 4 T cells,effector CD 4 T cells,effector memory CD 8 T cells and type 2 T helper cytokines were significantly elevated in patients with Brucella.Flow cytometry analysis identified higher proportions of Th2 and Th17 cells,and lower proportions of Th1 and Treg cells,in the peripheral blood in patients with brucellosis than healthy controls.The results of this study not only improve understanding of the immune response after Brucella infection but also provide directions for the diagnosis and treatment of brucellosis.