Adenosine augments IL-10 production by microglial cells through an A2B adenosine receptor-mediated process

J Immunol. 2012 Jan 1;188(1):445-53. doi: 10.4049/jimmunol.1101224. Epub 2011 Nov 23.

Abstract

Microglia are activated by pathogen-associated molecular patterns and produce proinflammatory cytokines, such as TNF-α, IL-6, and IL-12, and the anti-inflammatory cytokine IL-10. Adenosine is an endogenous purine nucleoside and a ligand of four G protein-coupled adenosine receptors (ARs), which are the A(1)AR, A(2A)AR, A(2B)AR, and A(3)AR. ARs have been shown to suppress TNF-α production by microglia, but their role in regulating IL-10 production has not been studied. In this study, we demonstrate that adenosine augments IL-10 production by activated murine microglia while suppressing the production of proinflammatory cytokines. Because the order of potency of selective AR agonists in inducing IL-10 production was NECA > IB-MECA > CCPA ≥ CGS21680, and the A(2B)AR antagonist MRS1754 prevented the effect of NECA, we conclude that the stimulatory effect of adenosine on IL-10 production is mediated by the A(2B)AR. Mechanistically, adenosine augmented IL-10 mRNA accumulation by a transcriptional process. Using mutant IL-10 promoter constructs we showed that a CREB-binding region in the promoter mediated the augmenting effect of adenosine on IL-10 transcription. Chromatin immunoprecipitation analysis demonstrated that adenosine induced CREB phosphorylation at the IL-10 promoter. Silencing CREB using lentivirally delivered short hairpin RNA blocked the enhancing effect of adenosine on IL-10 production, confirming a role for CREB in mediating the stimulatory effect of adenosine on IL-10 production. In addition, adenosine augmented IL-10 production by stimulating p38 MAPK. Collectively, our results establish that A(2B)ARs augment IL-10 production by activated murine microglia.

Publication types

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

MeSH terms

  • Acetamides / pharmacology
  • Adenosine / immunology*
  • Adenosine / pharmacology
  • Adenosine A2 Receptor Agonists / immunology
  • Adenosine A2 Receptor Agonists / pharmacology
  • Adenosine A2 Receptor Antagonists / pharmacology
  • Analgesics / immunology
  • Analgesics / pharmacology
  • Animals
  • CREB-Binding Protein / immunology
  • CREB-Binding Protein / metabolism
  • Cell Line
  • Interleukin-10 / biosynthesis
  • Interleukin-10 / immunology*
  • MAP Kinase Signaling System / drug effects
  • MAP Kinase Signaling System / immunology*
  • Mice
  • Microglia / cytology
  • Microglia / immunology*
  • Microglia / metabolism
  • Nerve Tissue Proteins / agonists
  • Nerve Tissue Proteins / antagonists & inhibitors
  • Nerve Tissue Proteins / immunology*
  • Nerve Tissue Proteins / metabolism
  • Phosphorylation / drug effects
  • Phosphorylation / immunology
  • Promoter Regions, Genetic / immunology
  • Purines / pharmacology
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / immunology
  • Receptor, Adenosine A2B / immunology*
  • Receptor, Adenosine A2B / metabolism
  • Transcription, Genetic / drug effects
  • Transcription, Genetic / immunology
  • p38 Mitogen-Activated Protein Kinases / immunology
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Acetamides
  • Adenosine A2 Receptor Agonists
  • Adenosine A2 Receptor Antagonists
  • Analgesics
  • IL10 protein, mouse
  • N-(4-cyanophenyl)-2-(4-(2,3,6,7-tetrahydro-2,6-dioxo-1,3-dipropyl-1H-purin-8-yl)-phenoxy)acetamide
  • Nerve Tissue Proteins
  • Purines
  • RNA, Messenger
  • Receptor, Adenosine A2B
  • Interleukin-10
  • CREB-Binding Protein
  • Crebbp protein, mouse
  • p38 Mitogen-Activated Protein Kinases
  • Adenosine