Negroni Anna, Stronati Laura, Pierdomenico Maria, Tirindelli Donatella, Di Nardo Giovanni, Mancini Valentina, Maiella Giulia, Cucchiara Salvatore
Section of Toxicology and Biomedical Sciences, University of Rome, Sapienza University Hospital Umberto I, Rome, Italy.
Inflamm Bowel Dis. 2009 Aug;15(8):1145-54. doi: 10.1002/ibd.20907.
NOD2 is an intracellular protein involved in host recognition of specific bacterial molecules and is genetically associated with several inflammatory diseases, including Crohn's disease (CD). NOD2 stimulation activates the transcription factor, NF-kappaB, through RIP2, a caspase-recruitment domain-containing kinase. NOD2/RIP2 signaling also mediates the activation of antimicrobial peptides such as human alpha-defensin 5 (HD-5) and human alpha-defensin 6 (HD-6), both produced by Paneth cells. The present study is aimed at describing the downstream events triggered specifically by NOD2 induction in order to demonstrate that the protein, other than overexpressed, is also physiologically associated with RIP2 and Erbin in the bioptic intestinal inflamed specimens of children affected by CD.
Fifteen children with CD and 10 children used as controls were entered in the study. Mucosal biopsy specimens were taken during endoscopy and mRNA and protein expressions were detected by using real-time polymerase chain reaction and Western blot.
NOD2 is able to form an immunocomplex with the kinase RIP2. As compared to controls, in the inflamed mucosa of patients both mRNA and protein expression levels of RIP2 are increased, and its active phosphorylated form is overexpressed.
In this study we provide for the first time ex vivo evidence of physiologically relevant protein interactions with NOD2, which are able to trigger the innate immune response in intestinal mucosal specimens of children with CD.
NOD2是一种细胞内蛋白,参与宿主对特定细菌分子的识别,并且在遗传上与包括克罗恩病(CD)在内的多种炎症性疾病相关。NOD2刺激通过RIP2(一种含半胱天冬酶招募结构域的激酶)激活转录因子NF-κB。NOD2/RIP2信号传导还介导抗菌肽的激活,如潘氏细胞产生的人α-防御素5(HD-5)和人α-防御素6(HD-6)。本研究旨在描述由NOD2诱导特异性触发的下游事件,以证明该蛋白除了过表达外,在受CD影响的儿童的活检肠道炎症标本中也在生理上与RIP2和Erbin相关。
15名患有CD的儿童和10名用作对照的儿童进入该研究。在内镜检查期间采集黏膜活检标本,并使用实时聚合酶链反应和蛋白质印迹法检测mRNA和蛋白质表达。
NOD2能够与激酶RIP2形成免疫复合物。与对照组相比,在患者的炎症黏膜中,RIP2的mRNA和蛋白质表达水平均升高,并且其活性磷酸化形式过表达。
在本研究中,我们首次提供了与NOD2发生生理相关蛋白相互作用的体外证据,这些相互作用能够在患有CD的儿童的肠道黏膜标本中触发先天性免疫反应。