Basic-Alimentary TractTNF-α and IFN-γ regulate the expression of the NOD2 (CARD15) gene in human intestinal epithelial cells☆,☆☆,★,★★,♢
Section snippets
Cell culture and transfection
Human epithelial HeLa S3 cells (ACC 161), intestinal epithelial HT-29 cells (ACC 299), and Caco-2 cells (ACC 169) were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). Two other colonic epithelial cell lines, SW620 and SW948 cells were kindly provided by Holger Kalthoff (UKK, Kiel, Germany). All cells were cultured in RPMI + 10% fetal calf serum. One day before transfection the cells were seeded at a density of 5 × 105 cells/2 mL on 6-well
NOD2 mRNA and protein expression is synergistically up-regulated in intestinal epithelial cell lines by TNF-α and IFN-γ
Different intestinal epithelial cell lines (HT-29, SW620, SW948) and HeLa S3 cells were stimulated for different durations with TNF-α (10 ng/mL), and total RNA was isolated as described. NOD2 complementary DNA was amplified as outlined and analyzed on an agarose gel (Figure 1A).
Conclusions
Epithelial cells are an integral part of the primary mechanical and immunological barrier against the invasion of microorganisms. The up-regulation of the NOD2 gene in IECs by TNF-α and IFN-γ could be part of the regulatory cascade of events during primary inflammatory processes. NOD2 has been shown to interact with RIP-like interacting CLARP kinase and overexpression of NOD2 in HEK293T cells led to activation of NF-κB.5 Interestingly, expression of NOD2 has been mainly presented in monocytic
References (49)
- et al.
Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3
Cell
(1997) - et al.
Human CARD4 protein is a novel CED-4/Apaf-1 cell death family member that activates NF-κB
J Biol Chem
(1999) - et al.
Nod1, an Apaf-1-like activator of caspase-9 and nuclear factor-κB
J Biol Chem
(1999) - et al.
Nod2, a Nod1/Apaf-1 family member that is restricted to monocytes and activates NF-κB
J Biol Chem
(2001) - et al.
Association between insertion mutation in NOD2 gene and Crohn's disease in German and British populations
Lancet
(2001) - et al.
The domains of death: evolution of the apoptosis machinery
Trends Biochem Sci
(1999) - et al.
Nods: a family of cytosolic proteins that regulate the host response to pathogens
Curr Opin Microbiol
(2002) - et al.
Signal transduction by tumor necrosis factor and its relatives
Trends Cell Biol
(2001) - et al.
Tumour necrosis factor alpha and interleukin 1beta in relapse of Crohn's disease
Lancet
(1999) - et al.
Mechanisms in failure of infliximab for Crohn's disease
Lancet
(2000)
Multiple nuclear factors interact with the immunoglobulin enhancer sequences
Cell
Nuclear factor κB is activated in macrophages and epithelial cells of inflamed intestinal mucosa
Gastroenterology
Induction of Nod2 in myelomonocytic and intestinal epithelial cells via nuclear factor-kappa B activation
J Biol Chem
Activation of nuclear factor-kappaB-dependent transcription by tumor necrosis factor-alpha is mediated through phosphorylation of RelA/p65 on serine 529
J Biol Chem
IKKbeta and phosphatidylinositol 3-kinase/Akt participate in non-pathogenic gram-negative enteric bacteria-induced RelA phosphorylation and NF-kappa B activation in both primary and intestinal epithelial cell lines
J Biol Chem
Mucosal flora in inflammatory bowel disease
Gastroenterology
The Caenorhabditis elegans cell death gene ced-4 encodes a novel protein and is expressed during the period of extensive programmed cell death
Development
The NOD: a signaling module that regulates apoptosis and host defense against pathogens
Oncogene
The NB-ARC domain: a novel signalling motif shared by plant resistance gene products and regulators of cell death in animals
Curr Biol
Plant pathogens and integrated defence responses to infection
Nature
A frameshift mutation in NOD2 associated with susceptibility to Crohn's disease
Nature
Association of NOD2 leucine-rich repeat variants with susceptibility to Crohn's disease
Nature
Interferons and their role in inflammation
Curr Pharm Des
Interferon expression in Crohn's disease patients: increased interferon-gamma and -alpha mRNA in the intestinal lamina propria mononuclear cells
J Interferon Res
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Address requests for reprints to: Dirk Seegert, Ph.D., Conaris Research Institute AG, Schauenburgerstr. 116, D-24118 Kiel, Germany. e-mail: [email protected]; fax: (49) 431-5606-822.
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Supported by grants from the “Deutsche Forschungsgemeinschaft” (SFB 415) and through a Competence Network from the German Ministry for Education and Research (BMBF) as well as from the National Genome Research Network.
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The authors thank the patients, the endoscopy staff, and the physicians (S. Nikolaus, U. Helwig, J. Grebe). The authors also thank G. Nunez for the pcDNA3NOD2wt plasmid. Ilka Woywod and Tanja Kaacksteen are gratefully acknowledged for their technical assistance.
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P. Rosenstiel and M. Fantini contributed equally to this work. D. Seegert and S. Schreiber share senior authorship.
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