In vivo single-cell high-dimensional mass cytometry analysis to track the interaction between Klebsiella pneumoniae and myeloid cells.
In vivo single-cell high-dimensional mass cytometry analysis to track the interaction between Klebsiella pneumoniae and myeloid cells.
复制标题
体内单细胞高维质谱流式分析追踪肺炎克雷伯菌与骨髓细胞之间的相互作用。
DOI：
10.1101/2023.12.14.571618
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Calderon-Gonzalez R
中科院分区：
文献类型：
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作者：
Calderon-Gonzalez R
体内单细胞方法已经改变了我们对组织中免疫群体的理解。将质谱的分辨率与以单细胞分辨率进行细胞分子的多重测量的能力相结合的质谱细胞术（CyTOF）已经能够解析免疫细胞亚群的多样性及其异质性功能。在这里，我们评估了CyTOF进一步免疫细胞的可行性，同时跟踪它们与细菌的相互作用，这种方法我们称之为Bac-CyTOF。本研究以肺炎克雷伯氏菌为病原菌，探讨肺炎小鼠模型的建立。利用Bac-CyTOF技术，我们获得了肺炎克雷伯菌强毒株感染小鼠后免疫细胞的图谱。图谱的特征在于肺泡和单核细胞衍生的巨噬细胞的数量减少。相反，中性粒细胞和炎症性单核细胞的特征是表达活性较低细胞标记物（例如免疫检查点PD-L1）的亚群增加。这些是被感染的细胞。我们发现VI型分泌系统（T6 SS）有助于塑造肺免疫景观。T6 SS通过将克雷伯氏菌从肺泡巨噬细胞转移到间质巨噬细胞并限制炎性单核细胞的感染来控制与单核细胞/巨噬细胞的相互作用。缺乏T6 SS导致表达活性细胞标志物的细胞增加，表达PD-L1的亚群减少。通过对克雷伯氏菌和鲍曼不动杆菌在体内存活能力有限的菌株进行检测，我们发现嗜中性粒细胞的高度募集、肺泡巨噬细胞和嗜酸性粒细胞的相对高水平以及嗜中性粒细胞特征性亚群的募集是小鼠清除感染的特征。我们利用Bac-CyTOF生成的知识平台来研究DNA传感器STING在克雷伯氏菌感染中的作用。sting-/-感染的小鼠呈现与清除感染一致的特征，包括PD-L1水平降低。STING缺失促进Klebsiella清除。
In vivo single-cell approaches have transformed our understanding of the immune populations in tissues. Mass cytometry (CyTOF), that combines the resolution of mass spectrometry with the ability to conduct multiplexed measurements of cell molecules at the single cell resolution, has enabled to resolve the diversity of immune cell subsets, and their heterogeneous functionality. Here we assess the feasibility of taking CyTOF one step further to immuno profile cells while tracking their interactions with bacteria, a method we term Bac-CyTOF. We focus on the pathogenKlebsiella pneumoniaeinterrogating the pneumonia mouse model. Using Bac-CyTOF, we unveil the atlas of immune cells of mice infected with aK.pneumoniaehypervirulent strain. The atlas is characterized by a decrease in the populations of alveolar and monocyte-derived macrophages. Conversely, neutrophils, and inflammatory monocytes are characterized by an increase in the subpopulations expressing markers of less active cells such as the immune checkpoint PD-L1. These are the cells infected. We show that the type VI secretion system (T6SS) contributes to shape the lung immune landscape. The T6SS governs the interaction with monocytes/macrophages by shiftingKlebsiellafrom alveolar macrophages to interstitial macrophages and limiting the infection of inflammatory monocytes. The lack of T6SS results in an increase of cells expressing markers of active cells, and a decrease in the subpopulations expressing PD-L1. By probingKlebsiella, andAcinetobacter baumanniistrains with limited ability to survive in vivo, we uncover that a heightened recruitment of neutrophils, and relative high levels of alveolar macrophages and eosinophils and the recruitment of a characteristic subpopulation of neutrophils are features of mice clearing infections. We leverage Bac-CyTOF-generated knowledge platform to investigate the role of the DNA sensor STING inKlebsiellainfections.sting-/-infected mice present features consistent with clearing the infection including the reduced levels of PD-L1. STING absence facilitatesKlebsiellaclearance.