PMID- 23918981 OWN - NLM STAT- MEDLINE DCOM- 20131106 LR - 20220129 IS - 1550-6606 (Electronic) IS - 0022-1767 (Print) IS - 0022-1767 (Linking) VI - 191 IP - 5 DP - 2013 Sep 1 TI - Two phases of inflammatory mediator production defined by the study of IRAK2 and IRAK1 knock-in mice. PG - 2717-30 LID - 10.4049/jimmunol.1203268 [doi] AB - The roles of IL-1R-associated kinase (IRAK)2 and IRAK1 in cytokine production were investigated using immune cells from knock-in mice expressing the TNFR-associated factor 6 (TRAF6) binding-defective mutant IRAK2[E525A] or the catalytically inactive IRAK1[D359A] mutant. In bone marrow-derived macrophages (BMDMs), the IRAK2-TRAF6 interaction was required for the late (2-8 h) but not the early phase (0-2 h) of il6 and tnfa mRNA production, and hence for IL-6 and TNF-alpha secretion by TLR agonists that signal via MyD88. Loss of the IRAK2-TRAF6 interaction had little effect on the MyD88-dependent production of anti-inflammatory molecules produced during the early phase, such as Dual Specificity Phosphatase 1, and a modest effect on IL-10 secretion. The LPS/TLR4-stimulated production of il6 and tnfa mRNA and IL-6 and TNF-alpha secretion was hardly affected, because the Toll/IL-1R domain-containing adapter-inducing IFN-beta (TRIF) signaling pathway was used instead of the IRAK2-TRAF6 interaction to sustain late-phase mRNA production. IRAK1 catalytic activity was not rate limiting for il6, tnfa, or il10 mRNA production or the secretion of these cytokines by BMDMs, but IFN-beta mRNA induction by TLR7 and TLR9 agonists was greatly delayed in plasmacytoid dendritic cells (pDCs) from IRAK1[D359A] mice. In contrast, IFN-beta mRNA production was little affected in pDCs from IRAK2[E525A] mice, but subsequent IFN-alpha mRNA production and IFN-alpha secretion were reduced. IFN-beta and IFN-alpha production were abolished in pDCs from IRAK1[D359A] x IRAK2[E525A] double knock-in mice. Our results establish that the IRAK2-TRAF6 interaction is rate limiting for the late, but not the early phase of cytokine production in BMDM and pDCs, and that the IRAK2-TRAF6 interaction is needed to sustain IkappaB-inducing kinase beta activity during prolonged activation of the MyD88 signaling network. [corrected] FAU - Pauls, Eduardo AU - Pauls E AD - Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, United Kingdom. AD - IrsiCaixa, Hospital Germans Trias i Pujol, Universitat Autonoma de Barcelona, Badalona 08916, Spain. FAU - Nanda, Sambit K AU - Nanda SK AD - Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, United Kingdom. FAU - Smith, Hilary AU - Smith H AD - Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, United Kingdom. FAU - Toth, Rachel AU - Toth R AD - Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, United Kingdom. FAU - Arthur, J Simon C AU - Arthur JSC AD - Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, United Kingdom. AD - Division of Cell Signaling and Immunology Unit, Sir James Black Centre, University of Dundee, DD1 5EH, United Kingdom. FAU - Cohen, Philip AU - Cohen P AD - Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, United Kingdom. LA - eng GR - MC_EX_G0800765/MRC_/Medical Research Council/United Kingdom GR - 100294/WT_/Wellcome Trust/United Kingdom GR - MC_EX_UU_G0800765/MRC_/Medical Research Council/United Kingdom GR - MR/K000985/1/MRC_/Medical Research Council/United Kingdom GR - WT_/Wellcome Trust/United Kingdom GR - MC_U127084348/MRC_/Medical Research Council/United Kingdom GR - MC_UU_12016/11/MRC_/Medical Research Council/United Kingdom PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20130805 PL - United States TA - J Immunol JT - Journal of immunology (Baltimore, Md. : 1950) JID - 2985117R RN - 0 (Cytokines) RN - 0 (TNF Receptor-Associated Factor 6) RN - EC 2.7.11.1 (Interleukin-1 Receptor-Associated Kinases) RN - EC 2.7.11.1 (Irak1 protein, mouse) RN - EC 2.7.11.10 (I-kappa B Kinase) RN - EC 2.7.11.10 (Ikbkb protein, mouse) SB - IM EIN - J Immunol. 2013 Nov 15;191(10):5317 MH - Animals MH - Bone Marrow Cells/immunology/metabolism MH - Cytokines/*biosynthesis MH - Dendritic Cells/immunology/*metabolism MH - Gene Knock-In Techniques MH - I-kappa B Kinase/immunology/metabolism MH - Immunoblotting MH - Immunoprecipitation MH - Inflammation/immunology/metabolism MH - Interleukin-1 Receptor-Associated Kinases/immunology/*metabolism MH - Macrophages/immunology/*metabolism MH - Mice MH - Real-Time Polymerase Chain Reaction MH - Signal Transduction/*immunology MH - TNF Receptor-Associated Factor 6/immunology/metabolism MH - Transfection PMC - PMC3849919 MID - EMS54158 OID - NLM: EMS54158 EDAT- 2013/08/07 06:00 MHDA- 2013/11/07 06:00 PMCR- 2014/03/01 CRDT- 2013/08/07 06:00 PHST- 2013/08/07 06:00 [entrez] PHST- 2013/08/07 06:00 [pubmed] PHST- 2013/11/07 06:00 [medline] PHST- 2014/03/01 00:00 [pmc-release] AID - 10.4049/jimmunol.1203268 [doi] PST - ppublish SO - J Immunol. 2013 Sep 1;191(5):2717-30. doi: 10.4049/jimmunol.1203268. Epub 2013 Aug 5.