基于单细胞测序技术探讨活动性肺结核CD8+T细胞功能耗竭亚群鉴定及分子调控网络注释

免疫细胞耗竭是影响结核病转归的重要因素，耗竭状态的CD8+T细胞可导致抑制性受体过表达、细胞因子信号通路失调及代谢适应性的改变。了解活动期肺结核患者CD8+T细胞不同亚群的发育和分类，建立CD8+T细胞功能耗竭与肺结核预后的关系亟待解决。本项目通过对活动期肺结核患者血液CD3+CD8+T细胞进行流式分选及单细胞转录组测序，明确活动期肺结核患者体内T细胞亚群的变化及分类，实现对功能耗竭CD8+T细胞亚群的鉴定；应用项目组前期建立的单细胞转录组Bi-cluster聚类技术及分子调控网络注释，解析CD8+T细胞亚群免疫耗竭的关键中枢节点分子，探讨其对免疫细胞耗竭的作用及机制，并最终建立活动性肺结核精准诊疗及预后评估的诊断标志物系统平台。本项目研究的特点是学科交叉合作与互补，从单细胞组学水平上解析结核感染的相关机制，构建结核感染CD8+T细胞耗竭分子网络模块并进行预后分析，为同类研究提供新思路。

Depletion of immune cells is an important factor affecting the outcome of tuberculosis. Depleted CD8 + T cells can lead to overexpression of inhibitory receptors, dysregulation of cytokine signaling pathways, and changes in metabolic adaptability. It is very urgent to understand the development and classification of different subsets of CD8 + T cells in patients with active tuberculosis, and establish the relationship between CD8 + T cell depletion and the prognosis of tuberculosis. In this project, through flow sorting and single cell transcriptome sequencing of blood CD3 + CD8 + T cells in patients with active tuberculosis, the changes and classification of T cell subsets in patients with active tuberculosis were clarified, and functionally exhausted CD8 + T cells were achieved identification of subgroups. Through the application of the single-cell transcriptome Bi-cluster clustering technology and molecular regulatory network annotations established earlier in the project team, we analyzed the key central node molecules of CD8 + T cell subtype immunodepletion and explored its role and mechanism on immune cell depletion. And finally establish a diagnostic marker cluster system platform for accurate diagnosis and prognosis evaluation of active pulmonary tuberculosis. The characteristics of this project research are cross-disciplinary cooperation and complementarity. Analyzing the related mechanism of tuberculosis infection from the level of single cell omics, constructing the molecular network module of tuberculosis infection CD8 + T cell depletion and prognostic analysis can provide new ideas for similar research.