Ghosh Susmita, Tuz Ali Ata, Stenzel Martin, Singh Vikramjeet, Richter Mathis, Soehnlein Oliver, Lange Emanuel, Heyer Robert, Cibir Zülal, Beer Alexander, Jung Marcel, Nagel Dennis, Hermann Dirk M, Hasenberg Anja, Grüneboom Anika, Sickmann Albert, Gunzer Matthias
Leibniz-Institut für Analytische Wissenschaften - ISAS - e.V., Dortmund, Germany.
Institute for Experimental Immunology and Imaging, University Hospital, University of Duisburg-Essen, Essen, Germany.
Mol Cell Proteomics. 2024 Nov;23(11):100858. doi: 10.1016/j.mcpro.2024.100858. Epub 2024 Oct 11.
Neutrophils are indispensable for defense against pathogens. Injured tissue-infiltrated neutrophils can establish a niche of chronic inflammation and promote degeneration. Studies investigated transcriptome of single-infiltrated neutrophils which could misinterpret molecular states of these post mitotic cells. However, neutrophil proteome characterization has been challenging due to low harvests from affected tissues. Here, we present a workflow to obtain proteome of 1000 murine and human tissue-infiltrated neutrophils. We generated spectral libraries containing ∼6200 mouse and ∼5300 human proteins from circulating neutrophils. 4800 mouse and 3400 human proteins were recovered from 1000 cells with 10-10 copies/cell. Neutrophils from stroke-affected mouse brains adapted to the glucose-deprived environment with increased mitochondrial activity and ROS-production, while cells invading inflamed human oral cavities increased phagocytosis and granule release. We provide an extensive protein repository for resting human and mouse neutrophils, identify proteins lost in low input samples, thus enabling the proteomic characterization of limited tissue-infiltrated neutrophils.
中性粒细胞对于抵御病原体至关重要。受伤组织中浸润的中性粒细胞可建立慢性炎症微环境并促进组织退化。此前的研究调查了单个浸润中性粒细胞的转录组，但这可能会误解这些终末分化细胞的分子状态。然而，由于从受影响组织中收获的细胞数量较少，中性粒细胞蛋白质组的表征一直具有挑战性。在此，我们展示了一种获取1000个小鼠和人类组织浸润中性粒细胞蛋白质组的工作流程。我们从循环中性粒细胞中生成了包含约6200种小鼠蛋白和约5300种人类蛋白的光谱库。从1000个细胞中回收了4800种小鼠蛋白和3400种人类蛋白，每个细胞有10-10个拷贝。中风小鼠大脑中浸润的中性粒细胞通过增加线粒体活性和活性氧生成来适应葡萄糖剥夺环境，而侵入人类发炎口腔的细胞则增加了吞噬作用和颗粒释放。我们为静息的人类和小鼠中性粒细胞提供了一个丰富的蛋白质库，鉴定了低输入样本中丢失的蛋白质，从而能够对有限的组织浸润中性粒细胞进行蛋白质组学表征。
