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Proteomic characterization of 1,000 human and murine neutrophils freshly isolated from blood and sites of sterile inflammation

ABSTRACT: Neutrophils, crucial player in inflammation, present a significant challenge in comprehensive molecular characterization due to limited availability (~1,000 cells) in inflamed sites like steady state infiltration. Prior proteomic studies typically required millions of neutrophils, posing difficulties when dealing with limited samples. This study introduces a ‘low-cell proteome’ workflow for neutrophils, integrating S-trap micro column-based digestion with DIA PASEF to investigate their functional dynamics within individual inflamed site.This method enable protein profiling of 1,000 isoated resting circulatory neutrophils with broad dynamic range in a reproducible manner.Furthermore, we applied this integrated approach to investigate the change in neutrophil proteome under different physiological and pathological conditions, including brain infiltration following stroke in mice and transmigration to the oral cavity post-treatment with tabasco in humans. Our comprehensive proteomic analysis provides insights into the distinct behavioral responses of neutrophils in these inflammatory contexts compared to their circulating counterparts.

INSTRUMENT(S): timsTOF

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Brain, Mouth, Neutrophil, Blood

DISEASE(S): Brain Ischemia

SUBMITTER: Susmita Ghosh

LAB HEAD: Albert Sickmann

PROVIDER: PXD050682 | Pride | 2024-10-11

REPOSITORIES: Pride

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			Action	DRS
	Activated_neutrophils_result.zip	Other
	Human_Mouthwash.zip	Other
	Human_blood_neutrophil.zip	Other
	Human_blood_neutrophil_result.zip	Other
	Human_neutrophil_library.kit	Other

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