基于单细胞核转录组测序的多发性大动脉炎外周血单个核细胞图谱分析  

Analysis of peripheral blood mononuclear cells in Takayasu arteritisbased on single cell nuclear transcriptome sequencing

作　　者：何川 丁洋[1] 万圣云[1] He Chuan;Ding Yang;Wan Shengyun(Dept of Vascular and Gastrointestinal Surgery,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601;Dept of Intervention,The Second Affiliated Hospital of Wannan Medical College,Wuhu 241000)

机构地区：[1]安徽医科大学第二附属医院血管与胃肠外科,合肥230601 [2]皖南医学院第二附属医院介入科,芜湖241000

出　　处：《安徽医科大学学报》2024年第6期1068-1075,共8页Acta Universitatis Medicinalis Anhui

基　　金：安徽高校自然科学研究项目重点项目(编号:2022AHO50764);安徽医科大学校科研基金项目临床项目(编号:2022xkj187)。

摘　　要：目的基于单细胞核转录组测序(snRNA-seq)技术描绘多发性大动脉炎(TAK)外周血单个核细胞(PBMC)基因和功能的异质性改变,阐述TAK可能的免疫机制。方法收集确诊为TAK患者的PBMC共6例为疾病组(TAK组),同期就诊的健康人群6例PBMC为正常组(NC组)。基于snRNA-seq工作流程建库测序,下机数据采用R 3.6.2软件进行主成分分析(PCA)及t-分布随机邻域嵌入(tSNE)分析等手段对两组PBMC样本池的单个核细胞的基因表达进行检测。结果TAK患者PBMC的免疫细胞的相对比例和含量都发生了改变;高克隆TCR可高度同源性匹配整合素beta-3的氨基酸序列;T细胞介导的干扰素γ和TNFα通路显著上调;TNFRSF13B+记忆B细胞比例明显升高;所有细胞类型中单核细胞所形成的的配体受体相互作用始终最多。结论TAK的发生发展过程中PBMC参与了复杂的调节,多种免疫炎症因子异常表达,其中单核细胞在PBMC的相互作用中占据了重要地位。Objective To characterize the genetic and functional heterogeneity of peripheral blood mononuclear cells(PBMC)in Takayasu arteritis(TAK)based on single cell nuclear transcriptome sequencing(snRNA-seq)technology and to elucidate the immune mechanisms that may be involved.Methods A total of 6 PBMC specimens were collected from patients who were diagnosed with TAK and had not received drug treatment recently and were admitted to the Second Affiliated Hospital of Anhui Medical University and the Second Affiliated Hospital of Wannan Medical College as the disease group(TAK group).During the same period,PBMC specimens were collected from patients who visited the hospital′s physical examination center.Six cases of PBMC in the population were in the normal group(NC group).Based on the snRNA-seq workflow,library construction and sequencing were performed,and R3.6.2 software was used to carry out PCA and tSNE analysis on the off-machine data from the two groups of PBMC sample pools for detection of gene expression in mononuclear cells.Results The relative proportion and content of immune cells in PBMC of TAK patients had changed;highly cloned TCR could match the amino acid sequence of integrin beta-3 with high homology;T cell-mediated interferon gamma and TNFαpathways were significantly up-regulated.The proportion of TNFRSF13B+memory B cells increased significantly.Monocytes consistently formed the largest number of ligand-receptor interactions of all cell types.Conclusion During the occurrence and development of TAK,PBMC are involved in complex regulation,and a variety of immune and inflammatory factors are abnormally expressed,among which monocytes play an important role in the interaction of PBMC.

关 键 词：单细胞核转录组测序 多发性大动脉炎 外周血单个核细胞 免疫 

分 类 号：R654.4[医药卫生—外科学]