Adenosine blocks IFN-γ-induced phosphorylation of STAT1 on serine 727 to reduce macrophage activation

Kimberly E. Barnholt, Rama S. Kota, Hnin Hnin Aung, John C Rutledge

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

Macrophages are activated by IFN-γ, a proinflammatory and proatherogenic cytokine that mediates its downstream effects primarily through STAT1. IFN-γ signaling induces phosphorylation of two STAT1 residues: Tyr701 (Y701), which facilitates dimerization, nuclear translocation, and DNA binding; and Ser727 (S727), which enables maximal STAT1 transcription activity. Immunosuppressive molecules such as adenosine in the cellular microenvironment can reduce macrophage inflammatory and atherogenic functions through receptor-mediated signaling pathways. We hypothesized that adenosine achieves these protective effects by interrupting IFN-γ signaling in activated macrophages. This investigation demonstrates that adding adenosine to IFN-γ-stimulated murine RAW 264.7 and human THP-1 macrophages results in unique modulation of STAT1 serine and tyrosine phosphorylation events. We show that adenosine inhibits IFN-γ-induced STAT1 S727 phosphorylation by >30% and phosphoserine-mediated transcriptional activity by 58% but has no effect on phosphorylation of Y701 or receptor-associated JAK tyrosine kinases. Inhibition of the adenosine A3 receptor with a subtype-specific antagonist (MRS 1191 in RAW 264.7 cells and MRS 1220 in THP-1 cells) reverses this adenosine suppressive effect on STAT1 phosphoserine status by 25-50%. Further, RAW 264.7 A3 receptor stimulation with Cl-IB-MECA reduces IFN-γ-induced STAT1 transcriptional activity by 45% and STAT1-dependent gene expression by up to 80%. These data suggest that A 3 receptor signaling is key to adenosine-mediated STAT1 modulation and anti-inflammatory action in IFN-γ-activated mouse and human macrophages. Because STAT1 plays a key role in IFN-γ-induced inflammation and foam cell transformation, a better understanding of the mechanisms underlying STAT1 deactivation by adenosine may improve preventative and therapeutic approaches to vascular disease.

Original languageEnglish (US)
Pages (from-to)6767-6777
Number of pages11
JournalJournal of Immunology
Volume183
Issue number10
DOIs
StatePublished - Nov 15 2009

ASJC Scopus subject areas

  • Immunology

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