Table 4_T-cell receptor and B-cell receptor repertoires profiling in pleural tuberculosis.xls

BackgroundTuberculosis (TB) is a leading cause of death worldwide from a single infectious agent. In China the most common extra-pulmonary TB (EPTB) is pleural tuberculosis (PLTB). An important clinical feature of PLTB is that the lymphocytes associated with TB will accumulate in the pleural fluid. The adaptive immune repertoires play important roles in Mycobacterium tuberculosis (Mtb) infection.
MethodsIn this study, 10 PLTB patients were enrolled, and their Peripheral Blood Mononuclear Cells(PBMCs) and Pleural Effusion Mononuclear Cells(PEMCs) were collected. After T cells were purified from PBMCs and PEMCs, high-throughput immunosequencing of the TCRβ chain (TRB), TCRγ chain(TRG), and B cell receptor(BCR) immunoglobulin heavy chain (IGH) were conducted on these samples.
ResultsThe TRB, TRG, and BCR IGH repertoires were characterized between the pleural effusion and blood in PLTB patients, and the shared clones were analyzed and collected. The binding activity of antibodies in plasma and pleural effusion to Mtb antigens was tested which indicates that different antibodies responses to Mtb antigens in plasma and pleural effusion in PLTB patients. Moreover, GLIPH2 was used to identify the specificity groups of TRB clusters and Mtb-specific TRB sequences were analyzed and collected by VJ mapping.
ConclusionWe characterize the adaptive immune repertoires and identify the shared clones and Mtb-specific clones in pleural effusion and blood in PLTB patients which can give important clues for TB diagnosis, treatment, and vaccine development.

背景
结核病（Tuberculosis, TB）是单一传染源导致全球死亡的首要病因。在中国，最常见的肺外结核（extra-pulmonary TB, EPTB）为胸膜结核（pleural tuberculosis, PLTB）。胸膜结核的一项重要临床特征为结核相关淋巴细胞会在胸腔积液中聚集。在结核分枝杆菌（Mycobacterium tuberculosis, Mtb）感染过程中，适应性免疫组库（adaptive immune repertoires）发挥着关键作用。

方法
本研究纳入10名胸膜结核患者，采集其外周血单个核细胞（Peripheral Blood Mononuclear Cells, PBMCs）与胸腔积液单个核细胞（Pleural Effusion Mononuclear Cells, PEMCs）。从PBMCs与PEMCs中纯化T细胞后，对上述样本开展T细胞受体β链（TCRβ chain, TRB）、T细胞受体γ链（TCRγ chain, TRG）以及B细胞受体（B cell receptor, BCR）免疫球蛋白重链（IGH）的高通量免疫测序。

结果
本研究对胸膜结核患者胸腔积液与血液中的TRB、TRG及BCR IGH免疫组库进行了特征解析，并对共享克隆型展开分析与收集。同时检测了血浆与胸腔积液中抗体针对结核分枝杆菌抗原的结合活性，结果显示胸膜结核患者血浆与胸腔积液内的抗体对结核分枝杆菌抗原的应答存在显著差异。此外，本研究使用GLIPH2工具鉴定TRB簇的特异性群组，并通过VJ作图分析与收集结核分枝杆菌特异性TRB序列。

结论
本研究对胸膜结核患者胸腔积液与血液中的适应性免疫组库进行了特征解析，鉴定出其中的共享克隆型与结核分枝杆菌特异性克隆型，该成果可为结核病的诊断、治疗以及疫苗研发提供重要线索。