Proteomics Analysis of Exosomes From Patients With Active Tuberculosis Reveals Infection Profiles and Potential Biomarkers.
Proteomics Analysis of Exosomes From Patients With Active Tuberculosis Reveals Infection Profiles and Potential Biomarkers.
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DOI：
10.3389/fmicb.2021.800807
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发表时间：
2021
影响因子：
5.2
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Zhang M;Xie Y;Li S;Ye X;Jiang Y;Tang L;Wang J
虽然结核病患者外周血清外周血中的外周血中的分枝杆菌蛋白已被鉴定，但外周血中外周血中的其他确切成分尚不清楚。本研究对活动性肺结核(ATB)患者血清外切体进行了全面的蛋白质组学分析。用纳米颗粒跟踪分析(NTA)、透射电子显微镜(TEM)和免疫印迹分析对ATB患者的外切体进行了表征。然后用无标记蛋白质组学对鉴定出的蛋白质组分进行定量，并通过生物信息学分析进行测定。从ATB血清外显体中鉴定出123个差异蛋白，并进行基因本体论(GO)分析。其中热休克蛋白70(HSP70)、CD81、主要组织相容性复合体-I(MHC-I)和肿瘤易感基因101(TSG101)存在于ATB和正常人的外体中。此外，在鉴定的胞外体蛋白中，脂多糖结合蛋白(LBP)在ATB外体中显著增加，而CD36和MHC-I显著降低。同时，ATB患者血清和外周血单个核细胞(PBMCs)中MHC-I表达下调，而CD36在血清中表达下调，而CD36在ATB患者PBMC中表达上调。Western blotting检测到结核分枝杆菌H37Ra感染巨噬细胞后，CD36在巨噬细胞中表达上调，而在H37Ra感染的巨噬细胞外体中CD36表达被抑制。本研究对ATB患者血清中的外切体蛋白质组进行了全面的描述，并揭示了与结核病感染相关的某些潜在的生物标志物。
Although mycobacterial proteins in exosomes from peripheral serum of patients with tuberculosis (TB) have been identified, other exact compositions of exosomes remain unknown. In the present study, a comprehensive proteomics analysis of serum exosomes derived from patients with active TB (ATB) was performed. Exosomes from patients with ATB were characterized using nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blotting analysis. Then identified protein components were quantified by label-free proteomics and were determined via bioinformatics analysis. A total of 123 differential proteins were identified in ATB serum exosomes and analyzed with Gene Ontology (GO) analysis. Among these proteins heat shock protein70 (HSP70), CD81, major histocompatibility complex-I (MHC-I ) and tumor susceptibility gene101 (TSG101) were present in exosomes of ATB and normal individuals confirmed via western blotting. In addition, among identified exosomal proteins lipopolysaccharide binding protein (LBP) increased significantly, but CD36 and MHC-I decreased significantly in ATB exosomes. Meanwhile, MHC-I was down-expressed in serum and peripheral blood mononuclear cells (PBMCs) of ATB, but interestingly CD36 was down-regulated in serum and up-expressed in PBMCs of ATB patients validated with ELISA and flow cytometry. CD36 was up-regulated by M. tuberculosis H37Ra infection in macrophages and suppressed in exosomes from H37Ra infected macrophages detected by western blotting. This study provided a comprehensive description of the exosome proteome in the serum of patients with ATB and revealed certain potential biomarkers associated with TB infection.