Open AccessMDP‐NOD2 stimulation induces HNP‐1 secretion, which contributes to NOD2 antibacterial function
Author(s) -
YamamotoFurusho Jesus K.,
Barnich Nicolas,
Hisamatsu Tadakazu,
Podolsky Daniel K.
Publication year - 2010
Publication title -
inflammatory bowel diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.932
H-Index - 146
eISSN - 1536-4844
pISSN - 1078-0998
DOI - 10.1002/ibd.21144
Subject(s) - nod2 , muramyl dipeptide , defensin , secretion , microbiology and biotechnology , biology , transfection , innate immune system , stimulation , chemistry , cell culture , immune system , biochemistry , immunology , peptide , endocrinology , genetics
Background: Human neutrophil peptide 1 (HNP‐1) is a defensin with antibacterial activity secreted by various cells as a component of the innate immune host defense. NOD2 is a cytoplasmic protein that recognizes bacterial derived muramyl dipeptide, and is involved in bacterial clearance. The aim of the present study was to investigate the relationship between antibacterial activity of NOD2 and HNP‐1 expression in epithelial cell lines. Methods: Gentamicin protection assay using Salmonella typhimurium was performed in Caco‐2 cells. The mRNA level was determined by quantitative reverse‐transcription polymerase chain reaction (RT‐PCR) and defensin expression was assessed by Western blot and enzyme‐linked immunosorbent assay (ELISA). Nuclear factor‐κB activation was assessed using pIV luciferase and Renilla plasmids. A NOD2 mutant was generated by site‐directed mutagenesis. Results: Among the defensins tested, only HNP‐1 expression is induced in colonic epithelial model HCT116 cells after MDP‐LD stimulation. HNP‐1 secretion is significantly increased after MDP‐LD stimulation in the cell supernatant of intestinal epithelial cells expressing endogenous NOD2, but not in cells that lack endogenous NOD2 expression. HNP‐1 is required for NOD2‐dependent NF‐κB activation after MDP‐LD stimulation since hnp‐1 siRNA transfection abrogated the response to MDP‐LD stimulation. The antibacterial function of NOD2 against S. typhimurium was impaired when expression of HNP‐1 was blocked by siRNA. Conclusions: HNP‐1 secretion depends on NOD2 stimulation by MDP‐LD and contributes to antibacterial activity in intestinal epithelial cells expressing endogenous NOD2, but not NOD2 3020insC mutant associated with increased susceptibility to Crohn's disease. (Inflamm Bowel Dis 2009)