Recent advances in the pathogenesis of Crohn's disease (CD) have suggested that an aberrant innate immune response initiates the cascade of events leading to T-cell activation and to disease development. NOD2 protein, which is mainly expressed by innate immunity cells, appears to play a key role against bacteria by triggering a host defense response through the activation of the transcriptor factor NF-κB and a consequent proinflammatory cytokine production. The present study was aimed at investigating the expression and activity of NOD2, NF-κB, and of 2 proinflammatory cytokines, TNFα and IL-1β, in mucosal biopsies of CD affected children compared to healthy controls.
In all, 22 children with active CD and 10 matched controls were entered in the study. mRNA and protein expressions were detected using reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blot; NF-κB binding activity was assessed by electromobility gel shift assay (EMSA).
NOD2 and IL-1β mRNAs were upregulated in CD children. Protein levels of NOD2, TNFα, and nuclear NF-κB, as well as the binding activity of NF-κB to a consensus DNA sequence, were significantly increased in inflamed mucosa of patients as compared to controls. Moreover, NF-κB activity was strongly upregulated in patients also when bound to the NOD2 promoter site. No difference was seen between patients and controls when NF-κB binding activity was determined in the uninflamed tissue.
This study suggests that altered mechanisms regulating NOD2 induction, NF-κB activation and cytokine production may contribute to dysregulate the innate immune response underlying pediatric CD.
1Section of Toxicology and Biomedical Sciences, Enea, Rome, Italy
2Department of Paediatrics, University of Rome, La Sapienza, University Hospital Umberto I, Rome, Italy
3Unità di Gastroenterologia, Ospedale IRCCS Casa Sollievo della Sofferenza San Giovanni Rotondo (Fg), Italy
*From the Professor of Pediatrics, Division of Pediatric Gastroenterology, Head, Department of Pediatrics, University of Rome “La Sapienza,” Viale Regina Elena, 324, 00161 Rome, Italy
Received 10 September 2007; Accepted 8 October 2007
Published online 18 December 2007 in Wiley Online Library (wileyonlinelibrary.com).