TNFR-associated factor 6 and TGF-β-activated kinase 1 control signals for a senescence response by an endosomal NK cell receptor

J Immunol. 2014 Jan 15;192(2):714-21. doi: 10.4049/jimmunol.1302384. Epub 2013 Dec 11.

Abstract

The endosomal innate receptor CD158d (killer cell Ig-like receptor 2DL4) induces cellular senescence in human NK cells in response to soluble ligand (HLA-G or agonist Ab). These senescent NK cells display a senescence-associated secretory phenotype, and their secretome promotes vascular remodeling and angiogenesis. To understand how CD158d initiates signaling for a senescence response, we mapped the region in its cytoplasmic tail that controls signaling. We identified a conserved TNFR-associated factor 6 (TRAF6) binding motif, which was required for CD158d-induced NF-κB activation and IL-8 secretion, TRAF6 association with CD158d, and TRAF6 recruitment to CD158d(+) endosomes in transfected cells. The adaptor TRAF6 is known to couple proximal signals from receptors such as endosomal TLRs and CD40 through the kinase TGF-β-activated kinase 1 (TAK1) for NF-κB-dependent proinflammatory responses. Small interfering RNA-mediated silencing of TRAF6 and TAK1, and inhibition of TAK1 blocked CD158d-dependent IL-8 secretion. Stimulation of primary, resting NK cells with soluble Ab to CD158d induced TRAF6 association with CD158d, induced TAK1 phosphorylation, and inhibition of TAK1 blocked the CD158d-dependent reprogramming of NK cells that produces the senescence-associated secretory phenotype signature. Our results reveal that a prototypic TLR and TNFR signaling pathway is used by a killer cell Ig-like receptor that promotes secretion of proinflammatory and proangiogenic mediators as part of a unique senescence phenotype in NK cells.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Amino Acid Motifs / genetics
  • Cell Line
  • Cellular Senescence / genetics*
  • Cytoplasm / genetics
  • Cytoplasm / metabolism
  • Endosomes / genetics
  • Endosomes / metabolism*
  • HEK293 Cells
  • Humans
  • Interleukin-8 / genetics
  • Interleukin-8 / metabolism
  • Killer Cells, Natural / metabolism*
  • MAP Kinase Kinase Kinases / genetics*
  • MAP Kinase Kinase Kinases / metabolism*
  • NF-kappa B / genetics
  • NF-kappa B / metabolism
  • Phosphorylation / genetics
  • Protein Binding / genetics
  • Receptors, KIR2DL4 / genetics
  • Receptors, KIR2DL4 / metabolism
  • Signal Transduction / genetics
  • TNF Receptor-Associated Factor 6 / genetics*
  • TNF Receptor-Associated Factor 6 / metabolism*
  • Toll-Like Receptors / genetics
  • Toll-Like Receptors / metabolism

Substances

  • Interleukin-8
  • KIR2DL4 protein, human
  • NF-kappa B
  • Receptors, KIR2DL4
  • TNF Receptor-Associated Factor 6
  • Toll-Like Receptors
  • MAP Kinase Kinase Kinases
  • MAP kinase kinase kinase 7