Xuefang Tao, Zhisong Xu, Hai Tian, Jingfeng He, Guowen Wang, Xuexia Tao
{"title":"基于串联质量标记分析的 EVs 差异蛋白质鉴定及 Treg 衍生 EVs 对 COPD 患者 T 淋巴细胞的影响","authors":"Xuefang Tao, Zhisong Xu, Hai Tian, Jingfeng He, Guowen Wang, Xuexia Tao","doi":"10.1186/s12931-024-02980-2","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Chronic obstructive pulmonary disease (COPD) is a widespread respiratory disease. This study examines extracellular vesicles (EVs) and proteins contained in EVs in COPD.</p><p><strong>Methods: </strong>Blood samples were collected from 40 COPD patients and 10 health controls. Cytokines including IFN-γ, TNF-α, IL-1β, IL-6, IL-8, and IL-17, were measured by ELISA. Small EVs samples were extracted from plasma and identified by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA), and Western blot. Protein components contained in EVs were analyzed by Tandem Mass Tags (TMT) to identify differential proteins. Treg-derived EV was extracted and added to isolated CD8<sup>+</sup>, Treg, and Th17 subsets to assess its effect on T-lymphocytes.</p><p><strong>Results: </strong>ELISA revealed higher levels of all cytokines and flow cytometry suggested a higher proportion of Treg and Th17 cells in COPD patients. After identification, TMT analysis identified 207 unique protein components, including five potential COPD biomarkers: BTRC, TRIM28, CD209, NCOA3, and SSR3. Flow cytometry revealed that Treg-derived EVs inhibited differentiation into CD8<sup>+</sup>, CD4<sup>+</sup>, and Th17 cells.</p><p><strong>Conclusion: </strong>The study shows that cytokines, T-lymphocyte subsets differences in COPD and Treg-derived EVs influence T-lymphocyte differentiation. Identified biomarkers may assist in understanding COPD pathogenesis, prognosis, and therapy. The study contributes to COPD biomarker research.</p>","PeriodicalId":49131,"journal":{"name":"Respiratory Research","volume":null,"pages":null},"PeriodicalIF":5.8000,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11439212/pdf/","citationCount":"0","resultStr":"{\"title\":\"Differential proteins from EVs identification based on tandem mass tags analysis and effect of Treg-derived EVs on T-lymphocytes in COPD patients.\",\"authors\":\"Xuefang Tao, Zhisong Xu, Hai Tian, Jingfeng He, Guowen Wang, Xuexia Tao\",\"doi\":\"10.1186/s12931-024-02980-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Chronic obstructive pulmonary disease (COPD) is a widespread respiratory disease. This study examines extracellular vesicles (EVs) and proteins contained in EVs in COPD.</p><p><strong>Methods: </strong>Blood samples were collected from 40 COPD patients and 10 health controls. Cytokines including IFN-γ, TNF-α, IL-1β, IL-6, IL-8, and IL-17, were measured by ELISA. Small EVs samples were extracted from plasma and identified by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA), and Western blot. Protein components contained in EVs were analyzed by Tandem Mass Tags (TMT) to identify differential proteins. Treg-derived EV was extracted and added to isolated CD8<sup>+</sup>, Treg, and Th17 subsets to assess its effect on T-lymphocytes.</p><p><strong>Results: </strong>ELISA revealed higher levels of all cytokines and flow cytometry suggested a higher proportion of Treg and Th17 cells in COPD patients. After identification, TMT analysis identified 207 unique protein components, including five potential COPD biomarkers: BTRC, TRIM28, CD209, NCOA3, and SSR3. Flow cytometry revealed that Treg-derived EVs inhibited differentiation into CD8<sup>+</sup>, CD4<sup>+</sup>, and Th17 cells.</p><p><strong>Conclusion: </strong>The study shows that cytokines, T-lymphocyte subsets differences in COPD and Treg-derived EVs influence T-lymphocyte differentiation. Identified biomarkers may assist in understanding COPD pathogenesis, prognosis, and therapy. The study contributes to COPD biomarker research.</p>\",\"PeriodicalId\":49131,\"journal\":{\"name\":\"Respiratory Research\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":5.8000,\"publicationDate\":\"2024-09-28\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11439212/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Respiratory Research\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1186/s12931-024-02980-2\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Respiratory Research","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s12931-024-02980-2","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0
摘要
背景:慢性阻塞性肺疾病(COPD)是一种广泛存在的呼吸系统疾病。本研究对慢性阻塞性肺病患者的细胞外囊泡 (EV) 和 EV 中所含的蛋白质进行了研究:方法:收集 40 名慢性阻塞性肺病患者和 10 名健康对照者的血液样本。采用 ELISA 法检测细胞因子,包括 IFN-γ、TNF-α、IL-1β、IL-6、IL-8 和 IL-17。从血浆中提取小的 EVs 样品,并通过透射电子显微镜(TEM)、纳米颗粒追踪分析(NTA)和 Western 印迹进行鉴定。利用串联质量标签(TMT)分析了EVs中的蛋白质成分,以确定不同的蛋白质。提取Treg衍生EV并将其加入分离的CD8+、Treg和Th17亚群,以评估其对T淋巴细胞的影响:结果:酶联免疫吸附试验(ELISA)显示慢性阻塞性肺病患者体内所有细胞因子的水平都较高,流式细胞术显示慢性阻塞性肺病患者体内 Treg 和 Th17 细胞的比例较高。经过鉴定,TMT 分析确定了 207 种独特的蛋白质成分,其中包括五种潜在的慢性阻塞性肺病生物标志物:BTRC、TRIM28、CD209、NCOA3 和 SSR3。流式细胞术显示,Treg衍生的EV抑制了CD8+、CD4+和Th17细胞的分化:研究表明,细胞因子、慢性阻塞性肺病的 T 淋巴细胞亚群差异以及 Treg 衍生的 EVs 会影响 T 淋巴细胞的分化。确定的生物标志物可能有助于了解慢性阻塞性肺病的发病机制、预后和治疗。该研究有助于慢性阻塞性肺病生物标志物的研究。
Differential proteins from EVs identification based on tandem mass tags analysis and effect of Treg-derived EVs on T-lymphocytes in COPD patients.
Background: Chronic obstructive pulmonary disease (COPD) is a widespread respiratory disease. This study examines extracellular vesicles (EVs) and proteins contained in EVs in COPD.
Methods: Blood samples were collected from 40 COPD patients and 10 health controls. Cytokines including IFN-γ, TNF-α, IL-1β, IL-6, IL-8, and IL-17, were measured by ELISA. Small EVs samples were extracted from plasma and identified by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA), and Western blot. Protein components contained in EVs were analyzed by Tandem Mass Tags (TMT) to identify differential proteins. Treg-derived EV was extracted and added to isolated CD8+, Treg, and Th17 subsets to assess its effect on T-lymphocytes.
Results: ELISA revealed higher levels of all cytokines and flow cytometry suggested a higher proportion of Treg and Th17 cells in COPD patients. After identification, TMT analysis identified 207 unique protein components, including five potential COPD biomarkers: BTRC, TRIM28, CD209, NCOA3, and SSR3. Flow cytometry revealed that Treg-derived EVs inhibited differentiation into CD8+, CD4+, and Th17 cells.
Conclusion: The study shows that cytokines, T-lymphocyte subsets differences in COPD and Treg-derived EVs influence T-lymphocyte differentiation. Identified biomarkers may assist in understanding COPD pathogenesis, prognosis, and therapy. The study contributes to COPD biomarker research.
期刊介绍:
Respiratory Research publishes high-quality clinical and basic research, review and commentary articles on all aspects of respiratory medicine and related diseases.
As the leading fully open access journal in the field, Respiratory Research provides an essential resource for pulmonologists, allergists, immunologists and other physicians, researchers, healthcare workers and medical students with worldwide dissemination of articles resulting in high visibility and generating international discussion.
Topics of specific interest include asthma, chronic obstructive pulmonary disease, cystic fibrosis, genetics, infectious diseases, interstitial lung diseases, lung development, lung tumors, occupational and environmental factors, pulmonary circulation, pulmonary pharmacology and therapeutics, respiratory immunology, respiratory physiology, and sleep-related respiratory problems.