中国防痨杂志2024，Vol.46Issue(4)：449-460,12.DOI:10.19982/j.issn.1000-6621.20240005
结核分枝杆菌潜伏感染者外周血单个核细胞转录组学研究
Transcriptome study on peripheral blood mononuclear cells of latent tuberculosis infection individuals
摘要
Abstract
Objective:The aim of the study is to explore the tuberculosis(TB)antigen specific transcriptome profile of latent TB infection(LTBI)individuals and identify the critical gene modules and pathways that may play significant roles in LTBI occurrence and development,by comparing the transcriptome results of peripheral blood mononuclear cells(PBMC)stimulated by TB-specific antigen of healthy controls(HC)and LTBI.Methods:Microarray test was used to uncover the transcriptome profile of PBMC stimulated by TB antigens in 4 cases of LTBI and 4 HC from Beijing Chest Hospital,Capital Medical University in December 2009,and further validated with qPCR to confirm the microarray data.Weighted gene co-expression network analysis(WGCNA)was used to identify the critical gene modules that were associated with LTBI.Geneontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were carried out based on the genes in the two modules significantly associated with LTBI.At the same time,GSEA-based KEGG analysis was also used in order to fully understand the transcriptome changes of PBMC after TB-specific antigen stimulation in the two groups.Immune infiltration analysis was also used to further confirm the important immune cells involving in LTBI occurrence.Protein-Protein Interaction(PPI)network analysis based on the STRING database was used to draw the differential gene interaction network diagram in the two modules.Results:A total of 393 differentially expressed genes were found between LTBI group and healthy group(fold change＞2 or＜0.5,P＜0.05),of which 112 were down-regulated and 281 were up-regulated.qPCR analysis of 10 differential genes confirmed that the gene expression pattern was consistent with that of the microarray data,and CAMK1G,IL2,SPINK1,SUCNR1,HCAR3,IL18BP,CHI3L1 and TNF were up-regulated,while RASGRP2 and TPM2 were down-regulated.According to the analysis of WGCNA,the blue module with positive correlation(cor=0.88,P=0.004)and the turquoise module with negative correlation(cor=-0.93,P=0.001)were the modules that most significantly associated with LTBI.The genes in the two modules were then subjected to KEGG enrichment analysis,meanwhile,GSEA analysis based on complete transcriptome was also performed.The results showed that chemokine signaling pathway(P＜0.001)and apoptosis signaling pathway(P=0.003)were significantly enriched by both analyses.The PPI network diagram of the differential genes in the two modules was constructed through the STRING database,and the top 10 hub genes in each module were obtained through the built-in plug-in,including PTPRC,CD40,IL10,IRF8,CCR1,CD80,TLR8,CLEC7A,CD83 and CD274 in the blue module,and TNF,ICAM1,TNFRSF4,HAVCR2,CD276,CCL4,CD33,IL1RN,NCF1 and FGR in the turquoise module.Immune infiltration analysis showed that innate immune cells,such as M1 macrophage(P=0.029)and M2 macrophage(P=0.001),were significantly concentrated in LTBI population.Conclusion:Significant differences in transcriptome of PBMC stimulated by TB-specific antigen are identified between LTBI and HC.These differential genes are mainly enriched in apoptosis signal pathway and chemokine signal pathway.Meanwhile,macrophage-mediated immune response plays an important role.These results collectively reveal the important role of innate immune response in LTBI occurrence.关键词
分枝杆菌,结核/分枝杆菌感染/转录因子/生物学标记/免疫Key words
Mycobacterium tuberculosis/Mycobacterium infections/Transcription factors/Biological markers/Immunity分类
医药卫生引用本文复制引用
尚雪恬,姬秀秀,杜博平,邢爱英,潘丽萍,董静,黄麦玲,孙琦,贾红彦,张蓝月,刘秋月,姚明旭,王颖超..结核分枝杆菌潜伏感染者外周血单个核细胞转录组学研究[J].中国防痨杂志,2024,46(4):449-460,12.基金项目
国家自然科学基金(82172279,82100011) （82172279,82100011）
北京市自然科学基金(L234055) National Natural Science Foundation(82172279,82100011) （L234055）
Beijing Natural Science Foundation(L234055) （L234055）
