Du Fengjiao, Deng Yunyun, Deng Ling, Du Boping, Xing Aiying, Tao Hong, Li Hua, Xie Li, Zhang Xinyong, Sun Tao, Li Hao
National Clinical Laboratory on Tuberculosis, Beijing Tuberculosis and Thoracic Tumor Institute, Beijing Chest Hospital, Capital Medical University, Beijing, China.
Hangzhou ImmuQuad Biotechnologies, Hangzhou, China.
Front Immunol. 2024 Nov 27;15:1473486. doi: 10.3389/fimmu.2024.1473486. eCollection 2024.
Tuberculosis (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 (Mtb) infection.
In 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.
The 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.
We 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.
结核病是全球由单一传染源导致死亡的主要原因。在中国,最常见的肺外结核(EPTB)是结核性胸膜炎(PLTB)。PLTB的一个重要临床特征是与结核相关的淋巴细胞会在胸腔积液中积聚。适应性免疫库在结核分枝杆菌(Mtb)感染中起重要作用。
本研究纳入了10例PLTB患者,收集其外周血单个核细胞(PBMCs)和胸腔积液单个核细胞(PEMCs)。从PBMCs和PEMCs中纯化T细胞后,对这些样本进行TCRβ链(TRB)、TCRγ链(TRG)和B细胞受体(BCR)免疫球蛋白重链(IGH)的高通量免疫测序。
对PLTB患者胸腔积液和血液中的TRB、TRG和BCR IGH免疫库进行了表征,并分析和收集了共享克隆。检测了血浆和胸腔积液中抗体与Mtb抗原的结合活性,这表明PLTB患者血浆和胸腔积液中对Mtb抗原的抗体反应不同。此外,使用GLIPH2识别TRB簇的特异性组,并通过VJ定位分析和收集Mtb特异性TRB序列。
我们对PLTB患者胸腔积液和血液中的适应性免疫库进行了表征,鉴定了共享克隆和Mtb特异性克隆,这可为结核病的诊断、治疗和疫苗开发提供重要线索。