Lyn kinase controls TLR4-dependent IKK and MAPK activation modulating the activity of TRAF-6/TAK-1 protein complex in mast cells.
Mast cells (MCs) control allergic reactions and contribute to protective innate immune responses through TLR4 activation. The tyrosine kinase Lyn is important to the high affinity IgE receptor (FcεRI) signal transduction system in MCs, but its role on the TLR4 signalling cascade is still elusive. Here, we characterized several TLR4-triggered responses in bone marrow-derived mast cells (BMMCs) from wild-type (WT) and Lyn(-/-) mice. We found that Lyn(-/-) MCs secreted lower amounts of TNF-α after LPS challenge when compared with WT cells. Lyn(-/-) BMMCs showed less MAPK, IκB phosphorylation and NF-κB nuclear translocation after TLR-4 triggering than WT cells. LPS-induced MAPK and inhibitor of IκB kinase (IKK) phosphorylation were importantly reduced in the absence of Lyn. A constitutive interaction between TNF receptor associated factor 6 (TRAF-6) and phosphorylated TGF-β-activated kinase (TAK-1) was observed in Lyn(-/-) BMMCs and this complex was insensitive to LPS addition. Lyn kinase was activated and associated to TRAF-6 shortly after LPS addition in WT MCs. Analyzing two local MC-dependent innate immune responses in vivo, we found that Lyn positively controls early TNF-α production and immune cell recruitment after an intraperitoneal injection of LPS. Our results indicate that Lyn plays a positive role in TLR4-induced production of TNF-α in MCs controlling the activity of the TRAF-6/TAK-1 protein complex.
Centro de Investigación y de Estudios Avanzados (Cinvestav), Mexico City, Mexico.
Martin
M
Avila
Adriana
A
Martinez-Juarez
Alfredo
A
Ibarra-Sanchez
Claudia
C
Gonzalez-Espinosa
D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D051550:I-kappa B Kinase / Q000378:metabolism; D007113:Immunity, Innate; D008070:Lipopolysaccharides / Q000008:administration & dosage; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism; D008407:Mast Cells / Q000276:immunology*; D051379:Mice; D018345:Mice, Knockout; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D015398:Signal Transduction / Q000276:immunology; D048029:TNF Receptor-Associated Factor 6 / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*; D000099151:MAP Kinase Kinase Kinase 7
D051550:I-kappa B Kinase; D008070:Lipopolysaccharides; D020930:MAP Kinase Kinase Kinases; D048029:TNF Receptor-Associated Factor 6; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D048051:p38 Mitogen-Activated Protein Kinases; D019061:src-Family Kinases; D000099151:MAP Kinase Kinase Kinase 7; C493487:Tlr4 protein, mouse; C060902:lyn protein-tyrosine kinase