Enhanced activity of NLRP3 inflammasome in peripheral blood cells of patients with active rheumatoid arthritis.
Enhanced activity of NLRP3 inflammasome in peripheral blood cells of patients with active rheumatoid arthritis.
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DOI：
10.1186/s13075-015-0775-2
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发表时间：
2015-09-19
影响因子：
4.9
通讯作者：
Sidiropoulos P
中科院分区：
文献类型：
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作者：
Choulaki C;Papadaki G;Repa A;Kampouraki E;Kambas K;Ritis K;Bertsias G;Boumpas DT;Sidiropoulos P
白细胞介素-1 β（IL-1β）是一种主要的炎性细胞因子，主要由先天性免疫细胞通过NLRP 3-炎性体激活产生。内在和外在的危险信号都可以激活NLRP 3。据报道，NLRP 3-炎性体组分的遗传变异影响类风湿性关节炎（RA）的易感性和严重性。我们试图评估活动性RA患者与健康个体相比NLRP 3-炎性体的活性。在基线和炎性小体活化时，通过免疫印迹法评估NLRP 3、ASC、半胱天冬酶-1原和活性半胱天冬酶-1、IL-1 β原和活性IL-1β的细胞内蛋白表达。在引发TLR 2（Pam（3）CysSK（4）、TLR 3（poly（I：C））或TLR 4（LPS）和ATP顺序处理后评估NLRP 3功能（IL-1β分泌）。我们使用半胱天冬酶抑制剂（casp-1、3/7和8）来评估它们对IL-1β成熟的贡献。所有实验均在全血细胞中进行。活动性RA患者（n = 11）表达更高的NLRP 3（p < 0.008）、ASC（p < 0.003）、活性半胱天冬酶-1（p < 0.02）和pro-IL-1β（p < 0.001）的基础细胞内水平。在用TLR 4（LPS）和ATP引发后，RA衍生的细胞提取物（n = 7）显示NLRP 3（p < 0.01）和活性半胱天冬酶-1（p < 0.001）的表达增加。RA患者（n = 20）用TLR 4（LPS）或TLR 3激动剂（poly（I：C））加ATP激活的全血细胞培养上清液中分泌的IL-1β高于对照组（n = 18）（两者均p < 0.02）。Caspase-1抑制显著降低所有刺激诱导的IL-1β分泌，而Caspase-8抑制仅影响TLR 4和TLR 3细胞引发。活动性RA患者在通过TLR 3和TLR 4而不是TLR 2刺激后，全血细胞中NLRP 3和NLRP 3介导的IL-1β分泌的表达增加。在这些患者中，IL-1β分泌似乎主要由caspase-1和caspase-8驱动。靶向NLRP 3或下游caspase可能有利于抑制RA中IL-1β的产生。
Interleukin-1β (IL-1β) is a major inflammatory cytokine, produced predominantly by innate immune cells through NLRP3-inflammasome activation. Both intrinsic and extrinsic danger signals may activate NLRP3. Genetic variations in NLRP3-inflammasome components have been reported to influence rheumatoid arthritis (RA) susceptibility and severity. We sought to assess the activity of NLRP3-inflammasome in patients with active RA compared to healthy individuals. Intracellular protein expression of NLRP3, ASC, pro- and active caspase-1, pro- and active IL-1β was assessed by immunoblotting both at baseline and upon inflammasome activation. NLRP3 function (IL-1β secretion) was assessed upon priming of TLR2 (Pam(3)CysSK(4), TLR3 (poly(I:C)) or TLR4 (LPS) and ATP sequential treatment. We used caspase inhibitors (casp-1, 3/7 and 8) to assess their contribution to IL-1β maturation. All experiments were performed in whole blood cells. Active RA patients (n = 11) expressed higher basal intracellular levels of NLRP3 (p < 0.008), ASC (p < 0.003), active caspase-1 (p < 0.02) and pro-IL-1β (p < 0.001). Upon priming with TLR4 (LPS) and ATP, RA-derived cell extracts (n = 7) displayed increased expression of NLRP3 (p < 0.01) and active caspase-1 (p < 0.001). Secreted IL-1β in culture supernatants from whole blood cells activated with TLR4 (LPS) or TLR3 agonist (poly(I:C)) plus ATP was higher in RA patients (n = 20) versus controls (n = 18) (p < 0.02 for both). Caspase-1 inhibition significantly reduced IL-1β secretion induced by all stimuli, whereas caspase-8 inhibition affected only TLR4 and TLR3 cell priming. Patients with active RA have increased expression of NLRP3 and NLRP3-mediated IL-1β secretion in whole blood cells upon stimulation via TLR3 and TLR4 but not TLR2. In these patients, IL-1β secretion seems to be predominately driven by caspase-1 and caspase-8. Targeting NLRP3 or downstream caspases may be of benefit in suppressing IL-1β production in RA.