Cold-inducible RNA-binding protein (CIRP) potentiates uric acid-induced IL-1ß production.

BACKGROUND: Gout is an autoinflammatory disease driven by interleukin-1 (IL-1) induction in response to uric acid crystals. IL-1ß production is dependent on inflammasome activation, which requires a priming signal, followed by an activating signal. The cold-inducible RNA-binding protein (CIRP) has been recently identified as a damage-associated molecular pattern (DAMP). In this study, we evaluated the roles of CIRP in monosodium urate (MSU)-mediated IL-1ß secretion using human neutrophils. METHODS: Human neutrophils were stimulated by MSU in the presence or absence of CIRP priming to determine NLRP3 inflammasome activation and subsequent caspase-1 activation and IL-1ß production. Cellular supernatants were analyzed by enzyme-linked immunosorbent assay (ELISA) to determine the presence of IL-1ß or caspase-1 (p20). The cellular supernatants and lysates were also analyzed by immunoblotting using anti-cleaved IL-1ß or anti-cleaved caspase-1 antibodies. RESULTS: Neither CIRP nor MSU stimulation alone induced sufficient IL-1ß secretion from neutrophils. However, MSU stimulation induced IL-1ß secretion from CIRP-primed neutrophils in a dose-dependent manner. This MSU-induced IL-1ß secretion from CIRP-primed neutrophils was accompanied by the induction of cleaved IL-1ß (p17), which was inhibited by the pretreatment of MCC950, a specific inhibitor for NLRP3. Furthermore, cleaved caspase-1 was induced in the cellular lysates of CIRP/MSU-treated neutrophils. Additionally, CIRP stimulation induced the protein expression of pro-IL-1ß in neutrophils. CONCLUSIONS: Our data indicate that CIRP, an endogenous stress molecule, triggers uric acid-induced mature IL-1ß induction as a priming stimulus for NLRP3 inflammasome in human neutrophils. We propose that CIRP acts as an important proinflammatory stimulant that primes and activates inflammasome and pro-IL-1ß processing in response to uric acid in innate immune cells.