Mouse Bone Marrow Neutrophil Extracellular Trap Proteomics by Microbial Stimuli

Neutrophils, the most abundant white blood cells in human circulation, play a crucial role in innate immunity. One of their key defense mechanisms is the formation of neutrophil extracellular traps (NETs), web-like structures composed of chromatin and antimicrobial proteins that help capture and neutralize pathogens. While previous studies have identified a limited set of NET-associated proteins, the comprehensive proteomic landscape of NETs induced by different stimuli remains largely unexplored. In this study, we used data-independent acquisition mass spectrometry to analyze the proteomic composition of NETs induced by five distinct stimuli: β-glucan, lipopolysaccharide, polyinosinic-polycytidylic acid sodium, resiquimod, and severe fever with thrombocytopenia syndrome bunyavirus. Across all conditions, we identified 5,868 NET-associated proteins, revealing significant stimulus-dependent differences in protein composition. Notably, differentially expressed proteins were detected in each condition, highlighting unique proteomic signatures that may reflect distinct immune responses. This dataset offers key insights into the proteomic diversity of NETs and their role in immune regulation, providing a foundation for future research on NET-mediated immunity in infectious and inflammatory diseases.