Cytokine production through PKC/p38 signaling pathways, not through JAK/STAT1 pathway, in mast cells stimulated with IFNgamma.
IFNgamma is strongly related to mast cell-associated diseases. There are many reports that IFNgamma inhibits mast cell degranulation. However, inflammatory cytokine production in mast cells stimulated with IFNgamma has not yet been clearly investigated. Therefore, we aimed to investigate the signaling pathways of cytokine production in mast cells stimulated with IFNgamma. Human mast cell line (HMC)-1 or mouse bone marrow-derived mast cells (BMMCs) were stimulated with IFNgamma (100 units) for time periods indicated. Expressions of proteins and mRNAs of cytokines were determined by ELISA and RT-PCR, respectively, activities of MAP kinases, PKC, JAK1/2, and STAT1 on tyrosine 701 and serine 727 by immunoblotting, the DNA-binding activity of the transcription factors by electrophoretic mobility shift assay. IFNgamma-stimulated mast cells showed increase in expressions of proteins and mRNAs of inflammatory cytokines, phosphorylations of MAP kinases, PKCalpha and betaI, JAK1/2, and STAT1 on tyrosine 701 and serine 727. JAK inhibitor or PKC inhibitors inhibited the phosphorylations of p38 kinase, STAT1 on serine 727, and activities of NF-kappaB and AP-1 compared to IFNgamma stimulation alone. These data suggest that IFNgamma-stimulated mast cells induce productions of inflammatory cytokines through PKC/p38/NF-kappaB and AP-1 pathways, not through classical JAK/STAT1 pathway, in both mast cells.
Seo
Jung Youn
JY
Department of Pharmacology and Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, 300 Chunchun-dong Jangan-ku, Suwon 440-746, Republic of Korea.
Kim
Dae Yong
DY
Lee
Yun Song
YS
Ro
Jai Youl
JY
D000818:Animals; D016207:Cytokines / Q000378:metabolism*; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D007249:Inflammation; D007371:Interferon-gamma / Q000378:metabolism*; D053613:Janus Kinase 1 / Q000378:metabolism*; D008407:Mast Cells / Q000201:enzymology*; D051379:Mice; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D050794:STAT1 Transcription Factor / Q000378:metabolism*; D018808:Transcription Factor AP-1 / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
D016207:Cytokines; D050794:STAT1 Transcription Factor; D018808:Transcription Factor AP-1; D007371:Interferon-gamma; D053613:Janus Kinase 1; D011493:Protein Kinase C; D048051:p38 Mitogen-Activated Protein Kinases