Related Experiment Video
Updated: Jul 7, 2026
Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine
Published on: May 21, 2012
Monocyte-derived dendritic cells from Crohn patients show differential NOD2/CARD15-dependent immune responses to
Valentina Salucci1, Monica Rimoldi, Chiara Penati
1Department of Experimental Oncology, European Institute of Oncology, Milan, Italy.
Background:
Three common mutations in the NOD2/CARD15 gene are strongly associated with Crohn's disease (CD). NOD2 is an intracellular receptor of muramyl dipeptide (MDP), a component of peptidoglycan present in the cell wall of gram-positive (G+) and gram-negative (G-) bacteria.
Methods:
We generated monocyte-derived dendritic cells (MoDCs) from CD patients mutated or not for CARD15 (n = 53) or from healthy donors (n = 12) and analyzed their activation in response to live Salmonella typhimurium as a model of pathogenic G- bacteria.
Results:
MoDCs carrying the L1007fs mutation, although phenotypically activated by bacteria, produced a significantly reduced amount of tested cytokines. MoDCs carrying R702W or compound G908R/R702W NOD2 mutations displayed an increased basal level of IL-8 release. After a bacterial encounter, these cells were phenotypically activated and produced levels of cytokines similar to healthy controls. Interestingly, although L1007fs/WT mutations conferred reduced production of cytokines, including IL-12, these cells were perfectly capable of inducing T-cell polarization toward the Th1 phenotype.
Conclusions:
NOD2 mutations affect the basal characteristics of MoDCs and their response to G- bacteria differently. MoDCs could be involved in CD onset because they have defects in releasing inflammatory cytokines and in polarizing T-cell responses.
Insights
NOD2 gene mutations impact immune cell responses to bacteria, potentially contributing to Crohn's disease (CD) development. Specific mutations alter cytokine release and T-cell polarization, revealing distinct cellular defects in CD patients.
Area of Science:
- Immunology
- Genetics
- Gastroenterology
Background:
- Three common NOD2/CARD15 gene mutations are linked to Crohn's disease (CD).
- NOD2 functions as an intracellular receptor for muramyl dipeptide (MDP), a bacterial cell wall component.
Purpose of the Study:
- To investigate how NOD2 mutations affect monocyte-derived dendritic cells (MoDCs) from CD patients.
- To analyze MoDC activation and cytokine production in response to Gram-negative bacteria.
Main Methods:
- Generated MoDCs from CD patients (with or without CARD15 mutations) and healthy donors.
- Stimulated MoDCs with live Salmonella typhimurium.
- Assessed MoDC activation, cytokine production (e.g., IL-8, IL-12), and T-cell polarization.
Main Results:
- L1007fs mutation led to reduced cytokine production but normal Th1 T-cell polarization.
- R702W or G908R/R702W mutations increased basal IL-8 release.
- Mutated MoDCs showed altered responses to Gram-negative bacteria compared to controls.
Conclusions:
- NOD2 mutations differentially affect MoDC characteristics and bacterial responses.
- MoDC defects in inflammatory cytokine release and T-cell polarization may contribute to Crohn's disease onset.
Related Concept Videos
01:25Inflammatory Bowel Disease III: Crohn's Disease
01:24T Cell Types and Functions
Th1 cells stimulate dendritic cells to express necessary co-stimulatory molecules on their surfaces for...
01:30Inflammatory Bowel Disease II: Crohn's Disease
Inflammatory bowel disease, commonly known as IBD, refers to a collection of disorders that lead to persistent inflammation of the gastrointestinal tract. The two types of IBD are ulcerative colitis, which impacts the colon, and Crohn's disease, which can involve any part of the gastrointestinal segment.
Crohn's disease
Crohn's disease is a chronic, systemic inflammatory bowel disease (IBD) that predominantly affects the gastrointestinal tract. It is marked by transmural...
01:31Defense Against Bacterial Pathogens
Phagocytes
Phagocytes are the frontline soldiers of the immune system. They include neutrophils and macrophages. Neutrophils are the most abundant type of white blood cell and are quickly mobilized to the site of infection. Macrophages are larger cells that patrol...