Signal transduction controls heterogeneous NF-κB dynamics and target gene expression through cytokine-specific refractory states

Nat Commun. 2016 Jul 6:7:12057. doi: 10.1038/ncomms12057.


Cells respond dynamically to pulsatile cytokine stimulation. Here we report that single, or well-spaced pulses of TNFα (>100 min apart) give a high probability of NF-κB activation. However, fewer cells respond to shorter pulse intervals (<100 min) suggesting a heterogeneous refractory state. This refractory state is established in the signal transduction network downstream of TNFR and upstream of IKK, and depends on the level of the NF-κB system negative feedback protein A20. If a second pulse within the refractory phase is IL-1β instead of TNFα, all of the cells respond. This suggests a mechanism by which two cytokines can synergistically activate an inflammatory response. Gene expression analyses show strong correlation between the cellular dynamic response and NF-κB-dependent target gene activation. These data suggest that refractory states in the NF-κB system constitute an inherent design motif of the inflammatory response and we suggest that this may avoid harmful homogenous cellular activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Feedback, Physiological
  • Gene Expression Regulation
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / immunology
  • Humans
  • I-kappa B Kinase / genetics
  • I-kappa B Kinase / immunology
  • Interleukin-1beta / pharmacology*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / immunology
  • NF-KappaB Inhibitor alpha / genetics*
  • NF-KappaB Inhibitor alpha / immunology
  • NF-kappa B / genetics*
  • NF-kappa B / immunology
  • Neurons
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / immunology
  • Receptors, Tumor Necrosis Factor, Type I / genetics*
  • Receptors, Tumor Necrosis Factor, Type I / immunology
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Red Fluorescent Protein
  • Signal Transduction / immunology*
  • Tumor Necrosis Factor alpha-Induced Protein 3 / antagonists & inhibitors
  • Tumor Necrosis Factor alpha-Induced Protein 3 / genetics
  • Tumor Necrosis Factor alpha-Induced Protein 3 / immunology
  • Tumor Necrosis Factor-alpha / pharmacology*


  • IL1B protein, human
  • Interleukin-1beta
  • Luminescent Proteins
  • NF-kappa B
  • RNA, Small Interfering
  • Receptors, Tumor Necrosis Factor, Type I
  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor-alpha
  • enhanced green fluorescent protein
  • NF-KappaB Inhibitor alpha
  • Green Fluorescent Proteins
  • I-kappa B Kinase
  • TNFAIP3 protein, human
  • Tumor Necrosis Factor alpha-Induced Protein 3