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Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine
Published on: May 21, 2012
Muramyl dipeptide-induced differential gene expression in NOD2 mutant and wild-type Crohn's disease patient-derived
Zuzana Zelinkova1, Astrid J van Beelen, Floor de Kort
1Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, The Netherlands. z.zelinkova@amc.uva.nl
Background:
Mutations in the gene encoding the nucleotide-binding oligomerization domain 2 (NOD2) protein are associated with Crohn's disease (CD), but the mechanism underlying this is not completely understood. To study the mechanism of CD resulting from NOD2 mutations, we analyzed NOD2-dependent whole-genome expression profiles of patient-derived antigen-presenting cells.
Patients And Methods:
Monocyte-derived dendritic cells (DCs) from CD carriers of double-dose NOD2 mutations, wild-type CD patients, and wild-type healthy volunteers were stimulated with the NOD2 ligand muramyl dipeptide. Whole-genome microarrays were used to assess the differential gene expression. The clustering of significantly changed genes was analyzed by online gene ontology mapping software.
Results:
In the DCs from the wild-type CD patient group, 683 genes were significantly changed, with most of the genes clustering in the pathways of inflammatory response. In addition, a significant number of genes clustered in the apoptosis regulation-related pathway. In the DCs from the healthy volunteer group, only 50 genes were significantly changed, predominantly those belonging to the response to pathogen pathway. Analysis of differentially expressed gene ontology pathways in the DCs from the NOD2 mutant CD patient group showed that the transcription of pathogen response genes was absent. In this group, 298 genes were significantly changed, predominantly clustering in the negative apoptosis regulation and cell organization and biogenesis pathways.
Conclusions:
Our results suggest that NOD2 mutations may result in perpetuation of mucosal inflammation through insufficient pathogen elimination. Further, these observations implicate a possible role of defective regulation of dendritic cell apoptosis in CD pathogenesis.
Insights
Mutations in the nucleotide-binding oligomerization domain 2 (NOD2) gene disrupt pathogen response and apoptosis regulation in Crohn's disease (CD) patients. This suggests NOD2 mutations perpetuate inflammation by impairing the immune system's ability to eliminate pathogens.
Area of Science:
- Immunology
- Genetics
- Gastroenterology
Background:
- Mutations in the nucleotide-binding oligomerization domain 2 (NOD2) gene are linked to Crohn's disease (CD), but the underlying mechanisms remain unclear.
- NOD2 plays a crucial role in innate immunity and regulating inflammatory responses.
Purpose of the Study:
- To investigate the functional consequences of NOD2 mutations on gene expression in immune cells.
- To elucidate the molecular mechanisms by which NOD2 mutations contribute to Crohn's disease pathogenesis.
Main Methods:
- Dendritic cells (DCs) were isolated from Crohn's disease patients with NOD2 mutations, wild-type CD patients, and healthy volunteers.
- Whole-genome microarrays were employed to analyze differential gene expression following stimulation with a NOD2 ligand.
- Gene ontology analysis was performed to identify enriched biological pathways.
Main Results:
- DCs from wild-type CD patients showed significant changes in inflammatory and apoptosis-related gene expression.
- DCs from healthy volunteers exhibited minimal gene expression changes, primarily related to pathogen response.
- DCs from NOD2 mutant CD patients displayed absent pathogen response gene transcription and altered apoptosis regulation pathways.
Conclusions:
- NOD2 mutations may lead to chronic mucosal inflammation due to impaired pathogen elimination.
- Defective regulation of dendritic cell apoptosis is implicated as a potential factor in Crohn's disease development.
