NLRP3 activation induces ASC-dependent programmed necrotic cell death, which leads to neutrophilic inflammation.
NLRP3 activation induces ASC-dependent programmed necrotic cell death, which leads to neutrophilic inflammation.
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DOI：
10.1038/cddis.2013.169
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发表时间：
2013-05-23
影响因子：
9
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中科院分区：
文献类型：
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NLR家族pyrin domain containing 3（NLRP 3）是一种胞质模式识别受体，其通过形成蛋白质复合物（炎性体）来调节先天性免疫应答。它导致产生促炎细胞因子产物，如白细胞介素1β（IL-1β）。我们和其他人证明，诱导活化的NLRP 3也诱导细胞死亡。然而，关于细胞死亡的特征和机制及其在炎症性疾病发病机制中的作用知之甚少。在这项研究中，我们建立了细胞系，其中NLRP 3诱导使用四环素诱导表达（Tet-on）系统的多西环素。使用该系统，NLRP 3突变体在cryopyrin-associated periodic syndrome（CAPS）患者中的表达足以诱导坏死细胞死亡，而无需脂多糖刺激或成熟IL-1β的产生。我们还发现，CA 074-Me，一种组织蛋白酶B抑制剂，在含有CARD的凋亡相关斑点样蛋白（ASC）寡聚化之前阻断细胞死亡，而Z-VAD-favour，一种泛半胱天冬酶抑制剂，在寡聚化之后阻断细胞死亡。通过小发夹RNA处理的ASC基因（Pycard）的沉默抑制NLRP 3突变体诱导的细胞死亡，但caspase-1基因（Casp 1）的沉默没有。总之，这些结果表明ASC对于NLRP 3介导的程序性坏死细胞死亡是不可或缺的，并且这种类型的细胞死亡不同于需要半胱天冬酶-1的“焦凋亡”。最后，我们在体内模型中证明，由活化的NLRP 3诱导的程序性坏死细胞死亡可引起中性粒细胞浸润，表明细胞死亡在CAPS患者皮肤病变的中性粒细胞浸润中可能起作用。
NLR family pyrin domain containing 3 (NLRP3) is a cytoplasmic pattern recognition receptor that regulates innate immune responses by forming a protein complex, the inflammasome. It leads to production of proinflammatory cytokine productions such as interleukin 1β (IL-1β). We and others demonstrated that an induction of activated NLRP3 also induced cell death. However, little is known about the characteristics and mechanisms of the cell death and its involvement in the pathogenesis of inflammatory conditions. In this study, we established cell lines in which NLRP3 was induced by doxycycline using a tetracycline-inducible expression (Tet-on) system. Using this system, the expression of NLRP3 mutants in cryopyrin-associated periodic syndrome (CAPS) patients was sufficient for the induction of necrotic cell death without lipopolysaccharide stimulation or generation of mature IL-1β. We also found that CA074-Me, a cathepsin B inhibitor, blocked cell death before oligomerization of apoptosis-associated speck-like protein containing a CARD (ASC), whereas Z-VAD-fmk, a pan-caspase inhibitor, blocked the cell death after the oligomerization. Silencing of the ASC gene (Pycard) by small hairpin RNA treatment inhibited the NLRP3 mutant-induced cell death, but silencing of the caspase-1 gene (Casp1) did not. Taken together, these results indicated that ASC was indispensable for NLRP3-mediated programmed necrotic cell death, and that this type of cell death was distinct from ‘pyroptosis', which requires caspase-1. Finally, we demonstrated in an in vivo model that the programmed necrotic cell death induced by activated NLRP3 could cause neutrophil infiltration, indicating a possible role of cell death in neutrophil infiltration of skin lesions in CAPS patients.
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影响因子：
64.8
作者：
Scaffidi, P;Misteli, T;Bianchi, ME
通讯作者：
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