BACKGROUND Tuberculosis (TB) continues to be a critical global health problem, which killed millions of lives each year. Certain circulating cell subsets are thought to differentially modulate the host immune response towards Mycobacterium tuberculosis (Mtb) infection, but the nature and function of these subsets is unclear. METHODS Peripheral blood mononuclear cells (PBMC) were isolated from healthy controls (HC), latent tuberculosis infection (LTBI) and active tuberculosis (TB) and then subjected to single-cell RNA sequencing (scRNA-seq) using 10 × Genomics platform. Unsupervised clustering of the cells based on the gene expression profiles using the Seurat package and passed to tSNE for clustering visualization. Flow cytometry was used to validate the subsets identified by scRNA-Seq. FINDINGS Cluster analysis based on differential gene expression revealed both known and novel markers for all main PBMC cell types and delineated 29 cell subsets. By comparing the scRNA-seq datasets from HC, LTBI and TB, we found that infection changes the frequency of immune-cell subsets in TB. Specifically, we observed gradual depletion of a natural killer (NK) cell subset (CD3-CD7+GZMB+) from HC, to LTBI and TB. We further verified that the depletion of CD3-CD7+GZMB+ subset in TB and found an increase in this subset frequency after anti-TB treatment. Finally, we confirmed that changes in this subset frequency can distinguish patients with TB from LTBI and HC. INTERPRETATION We propose that the frequency of CD3-CD7+GZMB+ in peripheral blood could be used as a novel biomarker for distinguishing TB from LTBI and HC. FUND: The study was supported by Natural Science Foundation of China (81770013, 81525016, 81772145, 81871255 and 91942315), National Science and Technology Major Project (2017ZX10201301), Science and Technology Project of Shenzhen (JCYJ20170412101048337) and Guangdong Provincial Key Laboratory of Regional Immunity and Diseases (2019B030301009). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

中文翻译：

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血液的单细胞转录组学揭示了结核病中自然杀伤细胞亚群的耗竭。

背景技术结核病(TB)仍然是一个严重的全球健康问题，每年夺去数百万人的生命。某些循环细胞亚群被认为可以差异调节宿主对结核分枝杆菌 (Mtb) 感染的免疫反应，但这些亚群的性质和功能尚不清楚。方法从健康对照（HC）、潜伏性结核感染（LTBI）和活动性结核（TB）中分离外周血单个核细胞（PBMC），然后使用10×Genomics平台进行单细胞RNA测序（scRNA-seq）。使用 Seurat 包基于基因表达谱对细胞进行无监督聚类，并传递给 tSNE 进行聚类可视化。流式细胞术用于验证 scRNA-Seq 识别的子集。结果 基于差异基因表达的聚类分析揭示了所有主要 PBMC 细胞类型的已知和新标记，并描绘了 29 个细胞亚群。通过比较来自 HC、LTBI 和 TB 的 scRNA-seq 数据集，我们发现感染改变了 TB 中免疫细胞亚群的频率。具体来说，我们观察到自然杀伤 (NK) 细胞亚群 (CD3-CD7+GZMB+) 从 HC 逐渐消耗到 LTBI 和 TB。我们进一步验证了 TB 中 CD3-CD7+GZMB+ 亚群的消耗，并发现抗结核治疗后该亚群频率增加。最后，我们证实该子集频率的变化可以将 TB 患者与 LTBI 和 HC 区分开来。解释 我们建议外周血中 CD3-CD7+GZMB+ 的频率可用作区分 TB 与 LTBI 和 HC 的新生物标志物。基金：该研究得到了国家自然科学基金（81770013、81525016、81772145、81871255和91942315）、国家科技重大专项（2017ZX10201301）、深圳市科技计划项目（JCYJ20170412101048337）和广东省区域免疫重点实验室的资助。疾病（2019B030301009）。资助者在研究设计、数据收集和分析、发表决定或手稿准备方面没有任何作用。