DUOX2-derived reactive oxygen species are effectors of NOD2-mediated antibacterial responses
DUOX2-derived reactive oxygen species are effectors of NOD2-mediated antibacterial responses
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DOI：
10.1242/jcs.050690
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发表时间：
2009-10-01
影响因子：
4
通讯作者：
Rosenstiel, Philip
中科院分区：
文献类型：
--
作者：
Lipinski, Simone;Till, Andreas;Rosenstiel, Philip
在许多真核生物中，杀微生物活性氧（ROS）的产生是先天免疫系统的关键保护成分。NOD（nucleotide oligomerisation domain containing protein，核苷酸寡聚化结构域）样受体（nucleotide oligomerisation domain containing protein，NLR）被认为是细胞内病原体或内源性危险信号的遗传学上古老的传感器。NOD 2识别细菌细胞壁组分胞壁酰二肽，通过丝氨酸-苏氨酸激酶RIP 2诱导的邻近信号传导导致NF κ B和MAPK活化。除了随后诱导细胞因子和抗微生物肽外，NOD 2还显示出直接的抗菌作用。使用基于荧光的ROS检测分析，我们证明了控制ROS生成作为一个不可分割的组成部分，NOD 2诱导的信号在上皮细胞。我们证明，NAD（P）H氧化酶家族成员DUOX 2参与NOD 2依赖性ROS的产生。免疫共沉淀和荧光显微镜显示DUOX 2与NOD 2在质膜上相互作用并共定位。此外，NOD 2和DUOX 2的同时过表达被发现导致针对细菌细胞侵入的合作保护，使用单核细胞增生李斯特菌感染模型。基于RNAi的研究表明，DUOX 2是NOD 2直接杀菌特性所必需的。我们的研究结果表明，一个新的作用的ROS作为效应分子的保护性细胞信号在响应一个确定的危险信号进行的哺乳动物细胞内NLR系统。
Generation of microbicidal reactive oxygen species (ROS) is a pivotal protective component of the innate immune system in many eukaryotes. NOD (nucleotide oligomerisation domain containing protein)-like receptors (NLRs) have been implicated as phylogenetically ancient sensors of intracellular pathogens or endogenous danger signals. NOD2 recognizes the bacterial cell wall component muramyldipeptide leading to NF kappa B and MAPK activation via induced proximity signalling through the serine-threonine kinase RIP2. In addition to the subsequent induction of cytokines and antimicrobial peptides, NOD2 has been shown also to exert a direct antibacterial effect. Using a fluorescence-based ROS detection assay we demonstrate controlled ROS generation as an integral component of NOD2-induced signalling in epithelial cells. We demonstrate that the NAD(P)H oxidase family member DUOX2 is involved in NOD2-dependent ROS production. Coimmunoprecipitation and fluorescence microscopy were used to show that DUOX2 interacts and colocalizes with NOD2 at the plasma membrane. Moreover, simultaneous overexpression of NOD2 and DUOX2 was found to result in cooperative protection against bacterial cytoinvasion using the Listeria monocytogenes infection model. RNAi-based studies revealed that DUOX2 is required for the direct bactericidal properties of NOD2. Our results demonstrate a new role of ROS as effector molecules of protective cellular signalling in response to a defined danger signal carried out by a mammalian intracellular NLR system.