[1]吾尔肯·居马巴依,许爱敏,邹小广.基于GEO/IEUOpenGWAS数据库及毒力因子ESAT-6/CFP-10免疫应答筛选与验证结核病核心炎症基因[J].现代检验医学杂志,2026,41(02):50-57.[doi:10.3969/j.issn.1671-7414.2026.02.009]
　WUERKEN Jumabayi,XU Aimin,ZOU Xiaoguang.Screening and Verification of Core Inflammatory Genes in Tuberculosis via GEO/IEU OpenGWAS Database Mining and ESAT-6/CFP-10 Virulence Factor Immune Response[J].Journal of Modern Laboratory Medicine,2026,41(02):50-57.[doi:10.3969/j.issn.1671-7414.2026.02.009]
点击复制

基于GEO/IEUOpenGWAS数据库及毒力因子ESAT-6/CFP-10免疫应答筛选与验证结核病核心炎症基因()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
50-57
栏目:
论著
出版日期:
2026-03-15

文章信息/Info

Title:
Screening and Verification of Core Inflammatory Genes in Tuberculosis via GEO/IEU OpenGWAS Database Mining and ESAT-6/CFP-10 Virulence Factor Immune Response
文章编号:
1671-7414（2026）02-050-08
作者:
吾尔肯·居马巴依1，许爱敏2，邹小广2
1.新疆医科大学第三临床医学院，乌鲁木齐 830000;2.喀什地区第一人民医院，新疆喀什 844000
Author(s):
WUERKEN Jumabayi1，XU Aimin2,3，ZOU Xiaoguang2,3
1.the Third Clinical Medical College of Xinjiang Medical University, Urumqi 830000, China;2.the First People'3.s Hospital of Kashi , Xinjiang Kashi 844000, China
关键词:
结核病; 早期分泌抗原6; 培养滤液蛋白10; 孟德尔随机化; 生物信息学
分类号:
R52；R446.11
DOI:
10.3969/j.issn.1671-7414.2026.02.009
文献标志码:
A
摘要:
目的?筛选并验证由结核分枝杆菌(MTB)毒力因子ESAT-6/CFP-10调控的关键基因,并阐明其在MTB感染中的作用,为开发新型结核病(TB)诊断标志物提供理论支持与候选靶点。方法基于GEO数据集GSE98461,整合差异表达分析与加权基因共表达网络分析(WGCNA),筛选靶基因并进行基因本体论(GO)功能注释与京都基因与基因组百科全书(KEGG)通路富集分析;利用Cytoscape构建蛋白互作(PPI)网络识别枢纽基因,分析免疫细胞浸润特征;进一步采用孟德尔随机化(MR)评估基因与TB的因果关联,并通过临床样本和RAW264.7巨噬细胞模型进行定量逆转录聚合酶链反应(RT-qPCR)验证。结果共筛选出414个靶基因,GO与KEGG分析显示其显著富集于免疫相关功能与通路;PPI网络分析进一步确定了IL-6、IL-10、PTGS2(COX-2)、IL-1A及CSF3(G-CSF)为核心枢纽基因;免疫浸润分析提示M1型巨噬细胞在免疫应答中发挥重要作用;MR分析表明IL-10与TB风险存在因果关系(P=0.0298);实验验证显示相较于对照组,ESAT-6/CFP-10刺激可显著上调巨噬细胞中基因IL-6、IL-10和PTGS2的mRNA表达水平,差异具有统计学意义(t=2.850~3.399,均P<0.05);相对于健康对照组,活动性结核患者外周血中基因IL-6与IL-10的mRNA水平也显著高表达,差异具有统计学意义(t=3.320、2.766,均P<0.01)。结论IL-6、IL-10和PTGS2在ESAT-6/CFP-10介导的结核免疫应答中具有核心作用,有望成为TB潜在检测与干预靶点。
Abstract:
Objective To identify key genes regulated by the Mycobacterium tuberculosis (MTB) virulence factors ESAT-6/CFP-10 and to elucidate their roles in MTB infection, thereby providing theoretical support and candidate targets for developing novel tuberculosis (TB) diagnostic biomarkers. Methods Based on GEO dataset GSE98461, differential expression analysis and Weighted Gene Co-expression Network Analysis (WGCNA) were integrated to screen candidate genes. Functional enrichment analysis was performed using Gene Ontology (GO) functional annotation and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Cytoscape was used to construct protein-protein interaction (PPI) networks to identify hub genes, and immune cell infiltration pattern. Mendelian randomization (MR) was employed to assess causal relationships between candidate genes and TB risk. Findings were validated using clinical samples and RAW264.7 macrophage model via quantitative reverse transcription PCR (RT-qPCR). Results 414 target genes were identified, with GO and KEGG analyses revealing signifi-cant enrichment in immune-related functions and pathways. PPI network analysis further identified IL-6, IL-10, PTGS2 (COX-2), IL-1A, and CSF3 (G-CSF) as hub genes. Immune infiltration analysis suggested M1 macrophages played a crucial role in the im-mune response. MR analysis indicated a causal relationship between IL-10 expression and TB risk (P=0.029 8). Experimental verification demonstrated that compared to the control group, ESAT-6/CFP-10 stimulation significantly upregulated the mRNA levels of IL-6, IL-10, and PTGS2 in macrophages (t=2.850~3.399, all P<0.05). Relative to the healthy controls, patients with active TB showed significantly higher mRNA levels of IL-6 and IL-10 in peripheral blood (t=3.320, 2.766, all P<0.01). Conclu-sions IL-6, IL-10, and PTGS2 play central roles in the ESAT-6/CFP-10-mediated immune response to tuberculosis, supporting their potential as targets for TB detection and intervention.

参考文献/References:

[1] 丁磊,徐俊驰,邱文娜,等.结核分枝杆菌耐药机制和治疗的最新研究进展[J].现代检验医学杂志, 2021, 36(2): 1-5, 48. DING L, XU J C, QIU W N, et al. Latest research prog-ress on the drug resistance mechanism and treatment of Mycobacterium tuberculosis[J]. Journal of Modern Laboratory Medicine, 2021, 36(2): 1-5, 48.
[2] NING J X, WANG R, PAN Y C, et al. Impact of tuber-culosis on median survival time and years of potential life lost in patients: a scoping review[J]. Journal of Clinical Tuberculosis and Other Mycobacterial Diseas-es, 2025, 41: 100568.
[3] SARAIVA M, VIEIRA P, O'GARRA A. Biology and therapeutic potential of interleukin-10[J]. Journal of Experimental Medicine, 2020, 217(1): e20190418.
[4] LIU S Y, XIAO G H, LI P X, et al. Plasma-based ul-trasensitive detection of Mycobacterium tuberculosis ESAT6/CFP10 fusion antigen using a CRISPR-driven aptamer fluorescence testing (CRAFT)[J]. Biosensors and Bioelectronics, 2025, 284: 117566.
[5] HUANG L, NAZAROVA E V, TAN S M, et al. Growth of Mycobacterium tuberculosis in vivo segregates with host macrophage metabolism and ontogeny[J]. Journal of Experimental Medicine, 2018, 215(4): 1135-1152.
[6] NIU Q, WANG M, LIU X S. The evolving landscape of IL-10, IL-22 and IL-26 in pleurisy especially in tubercu-lous pleurisy[J]. Respiratory Research, 2024, 25(1): 275.
[7] LIU K C, ZOU J, FAN H M, et al. Causal effects of gut microbiota on diabetic retinopathy: a Mendelian randomiza-tion study[J]. Frontiers in Immunology, 2022, 13: 930318.
[8] ZHANG H W, SU Y, HAI Y D, et al. The m6A regulatory gene YTHDF3 alleviates acute pancreatitis by modulating CD45RA+ resting treg cells: a novel immunomodulatory biomarker[J]. Medicine, 2025, 104(37): e44443.
[9] SEKULA P, DEL GRECO M F, PATTARO C, et al. Mendelian randomization as an approach to assess cau-sality using observational data[J]. Journal of the Ameri-can Society of Nephrology, 2016, 27(11): 3253-3265.
[10] 文维农,安祯祥,何远利,等.免疫细胞与肺结核的因果关系:一项孟德尔随机化研究[J].中国防痨杂志, 2024, 46(12):1519-1526. WEN W N, AN Z X, HE Y L, et al. The causal rela-tionship between immune cells and pulmonary tuber-culosis:a Mendelian randomization study[J]. Chinese Journal of Antituberculosis, 2024, 46(12): 1519-1526.
[11] MOUNIER N, KUTALIK Z. Bias correction for in-verse variance weighting Mendelian randomization[J]. Genetic epidemiology, 2023, 47(4): 314-331.
[12] 李苏亮,王哲,刘冰,等.基于孟德尔随机化研究体重指数与输血反应的因果关联[J].现代检验医学杂志, 2025, 40(6):149-153. LI S L, WANG Z, LIU B, et al. Study on the causal as-sociation between the body mass index and transfusion reactions based on Mendelian randomization[J]. Journal of Modern Laboratory Medicine, 2025, 40(6): 149-153.
[13] PENG Z L, XU C, YOU T X, et al. Depletion of Myco-bacterium tuberculosis transmembrane protein Rv3737 reduces pathogen survival and induces M1 macrophage polarization against tuberculosis[J]. Frontiers in Cellu-lar and Infection Microbiology, 2025, 15: 1592296.
[14] ZHU Z K, WANG H R, WANG X T, et al. The relation-ship between interleukin and ankylosing spondylitis[J]. Cytokine, 2025, 192: 156968.
[15] ALAM A, IMAM N, SIDDIQUI M F, et al. Human gene expression profiling identifies key therapeutic targets in tu-berculosis infection: a systematic network meta-analysis[J]. Infection Genetics and Evolution, 2021, 87: 104649.
[16] WEI J, GUO F Z, SONG Y M, et al. Transcriptional analy-sis of human peripheral blood mononuclear cells stimulated by Mycobacterium tuberculosis antigen[J]. Frontiers in Cellular and Infection Microbiology, 2023, 13: 1255905.
[17] 李建军,吴素方,白丰玺.基于Janus激酶/信号转导和转录激活因子信号通路的乙胺丁醇片对肺结核大鼠模型的作用机制[J].中国医学科学院学报, 2022, 44(4): 555-562. LI J J, WU S F, BAI F X. Action mechanism of etham-butol tablets on pulmonary tuberculosis rat model based on Janus kinase/signal transducer and activator of tran-scription signaling pathway [J]. Acta Academiae Medici-nae Sinicae, 2022,44(4):555-562.
[18] MUBEEN S A, VATS D, YADAV K, et al. CD14++CD16-classical monocyte subset secreting IL-1? and IL-10 is associated with ‘Tuberculosis Resisters’ phenotype[J]. Human Immunology, 2025, 86(2): 111240.
[19] MENSAH G I, AMPONSAH J A, ALAHAMAN N B, et al. Performance of eight serum cytokine/chemokine biomarkers in discriminating between active and latent tuberculosis infection in Ghana[J]. Frontiers in Immu-nology, 2025, 16: 1600712.
[20] SHEY M S, BALFOUR A, MASINA N, et al. Mycobac-terial-specific secretion of cytokines and chemokines in healthcare workers with apparent resistance to infection with Mycobacterium tuberculosis[J]. Frontiers in Immu-nology, 2023, 14: 1176615.
[21] LI F K, LUO J, XU H, et al. Early secreted antigenic target 6-kDa from Mycobacterium tuberculosis enhanced the protective innate immunity of macrophages partially via HIF1α[J]. Biochemical and Biophysical Research Communications, 2020, 522(1): 26-32.
[22] GUO Y D, LIU K H, YANG X T, et al. Multi-omics-based characterization of the influences of Mycobacterium tuberculosis virulence factors EsxB and PPE68 on host cells[J]. Archives of Microbiology, 2023, 205(6): 230.
[23] RUIZ-S?NCHEZ B P, CASTA?EDA-CASIMIRO J, CABRERA-RIVERA G L, et al. Differential activation of innate and adaptive lymphocytes during latent or ac-tive infection with Mycobacterium tuberculosis[J]. Mi-crobiology and Immunology, 2022, 66(10): 477-490.
[24] ABDALLA A E, LAMBERT N, DUAN X K, et al. Interleukin-10 family and tuberculosis: an old story re-newed[J]. International Journal of Biological Sciences, 2016, 12(6): 710-717.
[25] MOHAMMADNABI N, SHAMSEDDIN J, EMADI Mo bia, et al. Mycobacterium tuberculosis: the mechanism of pathogenicity, immune responses, and diagnostic challenges[J]. Journal of Clinical Laboratory Analysis, 2024, 38(23): e25122.
[26] CHEN Y C, LEE C P, HSIAO C C, et al. MicroR-NA-23a-3p down-regulation in active pulmonary tu-berculosis patients with high bacterial burden inhibits mononuclear cell function and phagocytosis through TLR4/TNF-α/TGF-β1/IL-10 signaling via targeting IRF1/SP1[J]. International Journal of Molecular Scienc-es, 2020, 21(22): 8587.
[27] FERREIRA C M, MICHELI C, BARREIRA-SILVA P, et al. IL-10 overexpression after BCG vaccination does not impair control of Mycobacterium tuberculosis infec-tion[J]. Frontiers in Immunology, 2022, 13: 946181.
[28] SILVA B D D S, TRENTINI M M, DA COSTA A C, et al. Different phenotypes of CD8+ T cells associated with bacterial load in active tuberculosis[J]. Immunology Let-ters, 2014, 160(1): 23-32.
[29] CLIFFORD V, TEBRUEGGE M, ZUFFEREY C, et al. Cytokine biomarkers for the diagnosis of tuberculosis infection and disease in adults in a low prevalence set-ting[J]. Tuberculosis, 2019, 114: 91-102.
[30] KIM S Y, PARK M S, KIM Y S, et al. The responses of multiple cytokines following incubation of whole blood from TB patients, latently infected individuals and controls with the TB antigens ESAT-6, CFP-10 and TB7.7[J]. Scan-dinavian Journal of Immunology, 2012, 76(6): 580-586.
[31] BAE M J, RYU S, KIM H J, et al. Mycobacterium tu-berculosis ESAT6 and CPF10 induce adenosine deam-inase 2 mRNA expression in monocyte-derived macro-phages[J]. Tuberculosis and Respiratory Diseases, 2017, 80(1): 77-82.
[32] AGRAWAL N. A comprehensive review on the advance-ments of dual COX-2/5-LOX inhibitors as anti-inflam-matory drugs[J]. Chemical Biology & Drug Design, 2025, 105(5): e70114.
[33] NING Y P, WANG W F, JORDAN P M, et al. Mycobac-terium tuberculosis-induced prostaglandin J2 and 15-de-oxy-prostaglandin J2 inhibit inflammatory signals in human M1 macrophages via a negative feedback loop[J]. Journal of Immunology, 2023, 210(10): 1564-1575.
[34] LI D Y, GAO C Y, ZHAO L, et al. Inflammatory response is modulated by lincRNACox2 via the NF-κB pathway in macrophages infected by Mycobacterium tuberculosis[J]. Molecular Medicine Reports, 2020, 21(6): 2513-2521.
[35] HA S H, CHOI H, PARK J Y, et al. Mycobacterium tuberculosis-secreted protein, ESAT-6, inhibits lipopoly-saccharide-induced MMP-9 expression and inflammation through NF-κB and MAPK signaling in RAW 264.7 macrophage cells[J]. Inflammation, 2020, 43(1): 54-65.
[36] A S K, BANSAL K, HOLLA S, et al. ESAT-6 induced COX-2 expression involves coordinated interplay be-tween PI3K and MAPK signaling[J]. Molecular Immu-nology, 2012, 49(4): 655-663.
[37] PANDEY R K, DAHIYA Y, SODHI A. Mycobacterium indicus pranii downregulates MMP-9 and iNOS through COX-2 dependent and TNF-α independent pathway in mouse peritoneal macrophages in vitro[J]. Microbes and Infection / Institut Pasteur, 2012, 14(4): 348-356.
[38] CARRANZA C, SARTILLO-MENDOZA L G, CARRE-TO-BINAGHI L E, et al. Exploring COX-2 inhibitors in tuberculosis: a whole-blood model approach for immune response and adjunt therapy evaluation[J]. Tuberculosis, 2023, 143: 102418.

备注/Memo

备注/Memo:
基金项目：国家自然科学基金（82260004）；新疆维吾尔自治区重点研发计划项目（2022B03032；2022B03032-3）
作者简介：吾尔肯?居马巴依（1999-），男，硕士在读，研究方向：结核病致病机制相关研究，E-mail：wuerken1007@163.com。
通讯作者：邹小广（1964-），男，博士，主任药师，研究方向：结核病致病机制及医院管理相关研究，E-mail：zxgkdyy1934@126.com。
更新日期/Last Update: 2026-03-15