TY - JOUR
T1 - IL-1β production is dependent on the activation of purinergic receptors and NLRP3 pathway in human macrophages
AU - Gicquel, Thomas
AU - Robert, Sacha
AU - Loyer, Pascal
AU - Victoni, Tatiana
AU - Bodin, Aude
AU - Ribault, Catherine
AU - Gleonnec, Florence
AU - Couillin, Isabelle
AU - Boichot, Elisabeth
AU - Lagente, Vincent
N1 - Publisher Copyright:
© FASEB.
PY - 2015/10/1
Y1 - 2015/10/1
N2 - The Nod-like receptor family protein 3 (NLRP3)-inflammasome pathway is known to be activated by danger signals such as monosodium urate (MSU). We investigated the role of P2 purinergic receptors in the activation of NLRP3-inflammasome pathway after MSU treatment of primary human monocyte-derived macrophages (MDMs). After initial stimulation with a low concentration of LPS (0.1 μg/ml), a 6 h treatment with MSU crystals (250, 500, and 1000 μg/ml) induced the MDMs to release IL-1β, IL-1α, and IL-6 in a dose-dependent manner. Moreover, the caspase 1 inhibitor Z-YVAD-FMK and the cathepsin B inhibitor CA-074Me reduced production of IL-1β in a dosedependent manner after LPS + MSU treatment. We used real-time reverse transcription-quantitative PCR to show that treatment with LPS and MSU (500 μg/ml) induced significantly greater expression of NLRP3 and IL-1β than after treatment with LPS. We also found that MSU treatment induced P2X purinergic receptor 7 (P2X7R) mRNA and protein expression. Furthermore, addition of the P2X7 purinergic receptor antagonist A-740003 significantly impeded IL-1β production and pro-IL-1β cleavage after treatment with LPS + MSU. Remarkably, RNA silencing of P2X7R (but not P2X4R) inhibited the release of IL-1β and other M1 macrophage cytokines (such as IL-1α, IL-6, and TNF-α) from MDMs stimulated with LPS + MSU. Taken as a whole, our results show that P2 purinergic receptors and the NLRP3 inflammasome pathway are involved in the secretion of IL-1β from MSU-stimulated human macrophages. This pathway may constitute a novel therapeutic target for controlling the inflammatory process in several associated pathologies.-Gicquel, T., Robert, S., Loyer, P., Victoni, T., Bodin, A., Ribault, C., Gleonnec, F., Couillin, I., Boichot,E.,Lagente,V.IL-1β production is dependent on the activation of purinergic receptors and NLRP3 pathway in human macrophages. FASEB J. 29, 4162-4173 (2015). www.fasebj.org.
AB - The Nod-like receptor family protein 3 (NLRP3)-inflammasome pathway is known to be activated by danger signals such as monosodium urate (MSU). We investigated the role of P2 purinergic receptors in the activation of NLRP3-inflammasome pathway after MSU treatment of primary human monocyte-derived macrophages (MDMs). After initial stimulation with a low concentration of LPS (0.1 μg/ml), a 6 h treatment with MSU crystals (250, 500, and 1000 μg/ml) induced the MDMs to release IL-1β, IL-1α, and IL-6 in a dose-dependent manner. Moreover, the caspase 1 inhibitor Z-YVAD-FMK and the cathepsin B inhibitor CA-074Me reduced production of IL-1β in a dosedependent manner after LPS + MSU treatment. We used real-time reverse transcription-quantitative PCR to show that treatment with LPS and MSU (500 μg/ml) induced significantly greater expression of NLRP3 and IL-1β than after treatment with LPS. We also found that MSU treatment induced P2X purinergic receptor 7 (P2X7R) mRNA and protein expression. Furthermore, addition of the P2X7 purinergic receptor antagonist A-740003 significantly impeded IL-1β production and pro-IL-1β cleavage after treatment with LPS + MSU. Remarkably, RNA silencing of P2X7R (but not P2X4R) inhibited the release of IL-1β and other M1 macrophage cytokines (such as IL-1α, IL-6, and TNF-α) from MDMs stimulated with LPS + MSU. Taken as a whole, our results show that P2 purinergic receptors and the NLRP3 inflammasome pathway are involved in the secretion of IL-1β from MSU-stimulated human macrophages. This pathway may constitute a novel therapeutic target for controlling the inflammatory process in several associated pathologies.-Gicquel, T., Robert, S., Loyer, P., Victoni, T., Bodin, A., Ribault, C., Gleonnec, F., Couillin, I., Boichot,E.,Lagente,V.IL-1β production is dependent on the activation of purinergic receptors and NLRP3 pathway in human macrophages. FASEB J. 29, 4162-4173 (2015). www.fasebj.org.
KW - Cytokines
KW - Nlrp3 inflammasome
KW - P2x7 receptor
KW - Uric acid
UR - http://www.scopus.com/inward/record.url?scp=84946945259&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84946945259&partnerID=8YFLogxK
U2 - 10.1096/fj.14-267393
DO - 10.1096/fj.14-267393
M3 - Article
C2 - 26116704
AN - SCOPUS:84946945259
SN - 0892-6638
VL - 29
SP - 4162
EP - 4173
JO - FASEB Journal
JF - FASEB Journal
IS - 10
ER -