Kyunghee Lee (Gyeongsang National University)

Eun Ju Jeong (Gyeongsang National University)

Hyuk-Kwon Kwon (Gyeongsang National University)

Kyunghee Lee (Gyeongsang National University)

Semin Lee (Gyeongsang National University)

Semin Park (Gyeongsang National University)

Dong-Won Bae (Gyeongsang National University)

Eun Ju Jeong (Gyeongsang National University)

Hyuk-Kwon Kwon (Gyeongsang National University)

The NLRP3 inflammasome is a key regulator of innate immune responses, but its effects on extracellular vesicle (EV) production and cargo composition remain unclear. Here, we characterized inflammatory EVs, termed infosomes, released from LPS- and nigericin-treated macrophages using label-free quantitative proteomics. NLRP3 activation increased EV abundance and protein content. Among 2,850 identified proteins, 1,314 were specifically detected in infosomes. Quantitative analysis identified 614 increased and 17 decreased proteins compared with control EVs. Infosomes were enriched in EV-associated, immune, cytoskeletal, membrane-trafficking, ribosomal, and RNA-binding proteins. Functional enrichment analysis highlighted immune responses, phagocytosis, endocytosis, translation, and RNA metabolism. NLRP3 knockout or MCC950 treatment reduced infosome protein loading, demonstrating NLRP3-dependent generation. Infosomes also induced inflammatory cytokine and chemokine expression in recipient macrophages and endothelial cells. These findings identify infosomes as proteome-enriched EVs that contribute to intercellular propagation of inflammation.

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