Lipopolysaccharide, IFN-gamma, and IFN-beta induce expression of the thiol-sensitive ART2.1 Ecto-ADP-ribosyltransferase in murine macrophages

J Immunol. 2007 Nov 1;179(9):6215-27. doi: 10.4049/jimmunol.179.9.6215.

Abstract

Nicotinamide adenosine dinucleotide (NAD) can act as a modulator of multiple immune and inflammatory responses when released into extracellular compartments. These actions of extracellular NAD are largely mediated by a family of mammalian ecto-ADP-ribosyltransferases (ARTs) that covalently modify target extracellular or cell surface proteins by transferring ADP-ribose to arginine or cysteine residues. In this study, we report that bone marrow-derived macrophages (BMDM) from BALB/c mice lack constitutive expression of any of the six murine ecto-ART subtypes, but selectively up-regulate ART2.1 in response to multiple proinflammatory mediators including agonists for TLR and type I and type II IFN. Stimulation of BMDM with LPS, IFN-beta, or IFN-gamma induced high expression of ART2.1, but not ART2.2, as a GPI-anchored cell surface ectoenzyme. ART2.1 expression in response to LPS was potentiated by inhibition of ERK1/2 signaling, but inhibited by blockade of the NF-kappaB, PI3K, and JAK-STAT pathways or the presence of neutralizing anti-IFN-beta. The catalytic function of the induced cell surface ART2.1 was strictly dependent on the presence of extracellular thiol-reducing cofactors, suggesting that in vivo activity of ART2.1-expressing macrophages may be potentiated in hypoxic or ischemic compartments. Consistent with the mutated art2a gene in C57BL/6 mice, LPS- or IFN-stimulated BMDM from this strain lacked expression of cell surface ART2 activity in the presence or absence of extracellular thiol reductants. Collectively, these studies identify ART2.1 as a new candidate for linking autocrine/paracrine activation of inflammatory macrophages to the release of NAD, a critical intracellular metabolite.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP Ribose Transferases / chemistry
  • ADP Ribose Transferases / genetics
  • ADP Ribose Transferases / metabolism*
  • Animals
  • Bone Marrow Cells / cytology
  • Cell Differentiation
  • Cell Membrane / drug effects
  • Cell Membrane / enzymology
  • Cells, Cultured
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Gene Expression Regulation, Enzymologic
  • Inflammation / enzymology
  • Inflammation / genetics
  • Interferon-beta / pharmacology*
  • Interferon-gamma / pharmacology*
  • Isoenzymes / genetics
  • Isoenzymes / metabolism
  • Lipopolysaccharides / pharmacology*
  • Macrophages / cytology
  • Macrophages / drug effects*
  • Macrophages / enzymology*
  • Mice
  • Models, Molecular
  • NF-kappa B / metabolism
  • Phosphatidylinositol 3-Kinases / metabolism
  • Protein Structure, Tertiary
  • Rats
  • Sensitivity and Specificity
  • Signal Transduction
  • Sulfhydryl Compounds / metabolism*

Substances

  • Isoenzymes
  • Lipopolysaccharides
  • NF-kappa B
  • Sulfhydryl Compounds
  • Interferon-beta
  • Interferon-gamma
  • ADP Ribose Transferases
  • Art2a protein, mouse
  • Phosphatidylinositol 3-Kinases
  • Extracellular Signal-Regulated MAP Kinases