基于串联质量标签分析的 EVs 差异蛋白鉴定及 COPD 患者 Treg 来源 EVs 对 T 淋巴细胞的影响。

Differential proteins from EVs identification based on tandem mass tags analysis and effect of Treg-derived EVs on T-lymphocytes in COPD patients.

机构信息

Department of Respiratory Medicine, The Affiliated Hospital of ShaoXing University, No. 999 Zhongxing South Road, Yuecheng District, Shaoxing, Zhejiang, 312000, China.

Department of Basic Medicine, Medical College, Shaoxing University, No. 900 Chengnan Avenue, Yuecheng District, Shaoxing, Zhejiang, 312000, China.

出版信息

Respir Res. 2024 Sep 28;25(1):349. doi: 10.1186/s12931-024-02980-2.

DOI:10.1186/s12931-024-02980-2

PMID:39342213

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11439212/

Abstract

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.

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。本研究探讨了 COPD 患者细胞外囊泡（EVs）及其所含蛋白质。

方法

采集 40 例 COPD 患者和 10 例健康对照者的血样。通过 ELISA 检测 IFN-γ、TNF-α、IL-1β、IL-6、IL-8 和 IL-17 等细胞因子。通过透射电子显微镜（TEM）、纳米颗粒跟踪分析（NTA）和 Western blot 提取并鉴定小 EVs 样本。通过串联质谱标签（TMT）分析 EV 中的蛋白成分，以鉴定差异蛋白。提取 Treg 衍生的 EV 并添加到分离的 CD8、Treg 和 Th17 亚群中，以评估其对 T 淋巴细胞的影响。

结果

ELISA 显示所有细胞因子水平升高，流式细胞术提示 COPD 患者 Treg 和 Th17 细胞比例较高。经鉴定，TMT 分析鉴定出 207 个独特的蛋白成分，包括 5 个潜在的 COPD 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 CD8、CD4 和 Th17 细胞的分化。