MIP-3α expression in macrophages is NOD dependent

Digestion. 2012;85(3):192-201. doi: 10.1159/000335423. Epub 2012 Jan 25.

Abstract

Background: The first identified susceptibility gene for Crohn's disease, NOD2, acts as a sensor for the bacterial-wall peptidoglycan fragment muramyl dipeptide (MDP) and activates the transcription factor nuclear factor-κB (NF-κB). Upon NF-κB activation, intestinal macrophages (IMACs) induce expression of macrophage inflammatory protein (MIP)-3α to attract memory T lymphocytes. We therefore investigated the influence of NOD2 ligation of IMAC differentiation and functional MIP-3α induction.

Methods: Human embryonal kidney HEK293 cells were transfected with NOD2 wild-type (NOD2(WT)) and the NOD2 SNP13 variant (NOD2(L1007fsinsC)) and stimulated with MDP. Recruitment of CD45R0+ and Th17 cells was determined by immunohistochemistry.

Results: Endogenous NOD2 stimulation was followed by a dose-dependent increase in MIP-3α secretion in MONO-MAC-6 (MM6) cells. MIP-3α mRNA was also significantly (*p < 0.05) induced in HEK293 transfected with NOD2(WT) via MDP ligation. In vivo cell-cell contacts between IMACs and CD45R0+ memory T cells as well as recruitment of Th17 cells in patients of NOD2 variants were unchanged as compared to wild-type patients.

Conclusion: Our data demonstrate a dose-dependent increase in MIP-3α secretion in the human myeloid cell line MM6 upon MDP. However, MIP-3α-driven recruitment of Th17 cells or CD45R0+ memory T lymphocytes is not affected in patients carrying heterozygous NOD2 variants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylmuramyl-Alanyl-Isoglutamine / pharmacology
  • Adolescent
  • Adult
  • Aged
  • Chemokine CCL20 / metabolism*
  • Crohn Disease / metabolism*
  • Crohn Disease / pathology
  • DNA Primers / chemistry
  • Dose-Response Relationship, Drug
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression
  • Genetic Predisposition to Disease
  • HEK293 Cells / drug effects
  • Humans
  • Immunohistochemistry
  • Intestinal Mucosa / pathology
  • Macrophages / metabolism*
  • Macrophages / pathology
  • Middle Aged
  • NF-kappa B / metabolism
  • Nod2 Signaling Adaptor Protein / physiology*
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • Transcriptional Activation
  • Transfection
  • Young Adult

Substances

  • CCL20 protein, human
  • Chemokine CCL20
  • DNA Primers
  • NF-kappa B
  • NOD2 protein, human
  • Nod2 Signaling Adaptor Protein
  • RNA, Messenger
  • Acetylmuramyl-Alanyl-Isoglutamine