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P2X7 receptor antagonism prevents IL-1β release from salivary epithelial cells and reduces inflammation in a mouse model of autoimmune exocrinopathy

Journal of Biological Chemistry. 2017; 
Mahmoud G. Khalafalla, Lucas T. Woods, Jean M. Camden, Aslam A. Khan, Kirsten H. Limesand, Michael J. Petris, Laurie Erb and Gary A. Weisman
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Recombinant Proteins NLRP3 in SMG cell lysates (500-1,000 µg protein/ml) was immunoprecipitated with protein A MagBeads (GenScript, Piscataway, NJ) pre-coated with mouse anti-mouse NLRP3 antibody (1:200 dilution; Cryo-2, Cat. No. AG-20B-0014-C100, Adipogen, San Diego, CA) by incubation for 1 h at room temperature. Immunoprecipitated proteins were solubilized using 40 µl of 1X SDS sample buffer (62.5 mM Tris-HCl at pH 6.8, 2% (w/v) SDS, 10% (v/v) glycerol, 50 mM DL-DTT and 0.01% (w/v) bromophenol blue) followed by heating at 95°C for 5 min. The samples were then subjected to SDS-PAGE on a 12% (w/v) gel (GenScript, Piscataway, NJ) and then transferred to nitrocellulose membranes for Western blot analysis. Get A Quote

摘要

Salivary gland inflammation is a hallmark of Sjögren’s syndrome (SS), a common autoimmune disease characterized by lymphocytic infiltration of the salivary gland and loss of saliva secretion, predominantly in women. The P2X7 receptor (P2X7R) is an ATP-gated non-selective cation channel that induces inflammatory responses in cells and tissues, including salivary gland epithelium. In immune cells, P2X7R activation induces the production of proinflammatory cytokines, including IL-1β and IL-18, by inducing the oligomerization of the multiprotein complex NLRP3-type inflammasome. Here, our results show that in primary mouse submandibular gland (SMG) epithelial cells, P2X7R activation also induces the assembly of ... More

关键词

ATP, inflammation, inflammasome, P2X7 receptor, salivary gland, Sjögren's syndrome