Ghosh, S.; Tuz, A. A.; Stenzel, M.; Singh, V.; Richter, M.; Soehnlein, O.; Lange, E.; Heyer, R.; Cibir, Z.; Beer, A.; Jung, M.; Nagel, D.; Hermann, D. M.; Hasenberg, A.; Grüneboom, A.; Sickmann, A.; Gunzer, M.
Abstract
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 1,000 murine and human tissue-infiltrated neutrophils. We generated spectral libraries containing ∼6,200 mouse and ∼5,300 human proteins from circulating neutrophils. 4,800 mouse and 3,400 human proteins were recovered from 1,000 cells with 102-108 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.
Keywords: Neutrophils; circulation; inflammation; low input proteomics; stroke; transmigration.