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通过高通量测序分析胸膜结核患者的T细胞受体库。

Profiling the T-cell receptor repertoire of patient with pleural tuberculosis by high-throughput sequencing.

作者信息

Li Dengrui, Gao Guanju, Li Zhihui, Sun Wei, Li Xiaoxia, Chen Ning, Sun Jingjing, Yang Yonghui

机构信息

Department of Pathology, Hebei Chest Hospital, 372 Shengli North Street, Shijiazhuang 050041, Hebei Province, China.

Department of Pathology, Hebei Chest Hospital, 372 Shengli North Street, Shijiazhuang 050041, Hebei Province, China.

出版信息

Immunol Lett. 2014 Nov;162(1 Pt A):170-80. doi: 10.1016/j.imlet.2014.08.012. Epub 2014 Aug 27.

DOI:10.1016/j.imlet.2014.08.012
PMID:25173045
Abstract

Pleural tuberculosis (PLTB), a major cause of morbidity and mortality, is the most common extrapulmonary manifestation of active Mycobacterium tuberculosis (Mtb) in developing countries. Gamma delta T-cell receptor (TCR) repertoire of peripheral blood mononuclear cells (PBMCs) and pleural effusion mononuclear cells (PEMCs) and beta TCR repertoire from peripheral blood mononuclear cells (PBMCs) have been reported. However, a detailed different characteristic of beta TCR repertoire of mononuclear cells isolated from peripheral blood and pleural fluid in the immune response to Mtb infection should be further revealed. The TCR β-chain (TRB) from PBMCs and PEMCs from an untreated pleural tuberculosis patient was sequenced by the Illumina sequencing platform. A total of 96,758 and 124,130 unique complementarity-determining region 3 (CDR3) sequences were identified at the nucleotide level, encoding 69,488 and 99,095 peptide sequences, respectively. TCR profiling showed that TRBV20-1 family and TRBV20-1/TRBJ1-5 gene combination had a dominant expression in PEMCs, but not in PBMCs. Expansive expression of common CDR3 clonotypes was observed in PEMCs. CDR3 spectratyping analysis showed that few TRBV families had a significantly skewed pattern, with one peak or a few prominent peaks in the PBMCs. By contrast, some TRBV families showed oligoclonal or clonal expansion in the PEMCs. Here, we firstly profiled the TRB repertoire differences of PBMCs and PEMCs from one PLTB patient using high-throughput sequencing. And this study may provide new insight for the detailed and efficient study of TCR repertoire of PEMCs in the future.

摘要

胸膜结核(PLTB)是发病和死亡的主要原因,是发展中国家活动性结核分枝杆菌(Mtb)最常见的肺外表现。外周血单个核细胞(PBMCs)和胸腔积液单个核细胞(PEMCs)的γδ T细胞受体(TCR)库以及外周血单个核细胞(PBMCs)的β TCR库已有报道。然而,在Mtb感染的免疫反应中,从外周血和胸腔积液中分离的单个核细胞β TCR库的详细差异特征仍有待进一步揭示。通过Illumina测序平台对一名未经治疗的胸膜结核患者的PBMCs和PEMCs中的TCR β链(TRB)进行测序。在核苷酸水平上共鉴定出96,758和124,130个独特的互补决定区3(CDR3)序列,分别编码69,488和99,095个肽序列。TCR分析表明,TRBV20-1家族和TRBV20-1/TRBJ1-5基因组合在PEMCs中呈优势表达,而在PBMCs中则不然。在PEMCs中观察到常见CDR3克隆型的扩增表达。CDR3光谱分型分析表明,PBMCs中很少有TRBV家族呈现明显的偏态模式,只有一个峰或几个突出的峰。相比之下,一些TRBV家族在PEMCs中显示出寡克隆或克隆性扩增。在此,我们首次使用高通量测序分析了一名PLTB患者的PBMCs和PEMCs的TRB库差异。本研究可能为未来详细、高效地研究PEMCs的TCR库提供新的见解。

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