Accumulation of glutathione disulfide mediates NF-κB activation during immune stimulation with CpG DNA

Jeffrey D. Kirsch, Ae-Kyung Yi, Douglas R. Spitz, Arthur M. Krieg

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Innate immune cells recognize pathogens by detecting molecular patterns that are distinct from those of the host. One such pattern is unmethylated CpG dinucleotides, which are common in bacterial DNA but not in vertebrate genomes. Macrophages respond to such CpG motifs in bacterial DNA or synthetic oligodeoxynucleotides (ODN) by inducing NF-κB and secreting proinflammatory cytokines, such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), but the mechanisms regulating this have been unclear. CpG ODN-stimulated cells produce reactive oxygen species (ROS) and have a decreased ratio of intracellular glutathione/glutathione disulfide (GSH/GSSG), indicating a shift to a more oxidized intracellular redox state. To determine whether this may play a role in mediating the CpG-induced macrophage activation, the GSH/GSSG redox state was manipulated in the murine macrophagelike cell line RAW264.7. Treatment of cells with BCNU to inhibit glutathione reductase (GR) enhanced the CpG-induced intracellular oxidation and decreased the GSH/GSSG, with increased activation of NF-κB and a doubling in the CpG-induced production of IL-6 and TNF-α. Experimental manipulation of the intracellular GSSG concentration during inhibition of cellular prooxidant production demonstrated that increased intracellular GSSG is a primary signal that is directly or indirectly required for CpG-induced NF-κB activation but is not in itself sufficient to trigger this in the absence of CpG ODN. These data suggest the existence of a second CpG-induced intracellular signal, independent of GSSG, mediating the activation of innate immunity by bacterial DNA.

Original languageEnglish (US)
Pages (from-to)327-340
Number of pages14
JournalAntisense and Nucleic Acid Drug Development
Volume12
Issue number5
DOIs
StatePublished - Oct 1 2002

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Glutathione Disulfide
Chemical activation
DNA
Bacterial DNA
Oligodeoxyribonucleotides
Macrophages
Oxidation-Reduction
Interleukin-6
Tumor Necrosis Factor-alpha
Cells
Carmustine
Macrophage Activation
Glutathione Reductase
Pathogens
Innate Immunity
Glutathione
Vertebrates
Reactive Oxygen Species
Genes
Genome

All Science Journal Classification (ASJC) codes

  • Genetics
  • Pharmacology

Cite this

Accumulation of glutathione disulfide mediates NF-κB activation during immune stimulation with CpG DNA. / Kirsch, Jeffrey D.; Yi, Ae-Kyung; Spitz, Douglas R.; Krieg, Arthur M.

In: Antisense and Nucleic Acid Drug Development, Vol. 12, No. 5, 01.10.2002, p. 327-340.

Research output: Contribution to journalArticle

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