Early and late events in Fc epsilon RI signal transduction in human cultured mast cells.
Protein tyrosine phosphorylation and other biochemical events have been shown to occur after cross-linking of Fc epsilonRI in rodent mast cells. To investigate the mechanism of Fc epsilonRI signal transduction in human mast cells, we used human cultured mast cells (HCMC) generated from cord blood cells in the presence of recombinant human stem cell factor and IL-6. We found that on cross-linking of Fc epsilonRI: 1) HCMC released histamine; 2) rapid tyrosine phosphorylation of multiple cellular substrates, including Syk, HS1, c-Cbl, ERK-1, and ERK-2, was observed; 3) intracellular Ca2+ and inositol phosphate production were increased within the first minute after Fc epsilonRI cross-linking; and 4) genistein, a tyrosine kinase inhibitor, inhibited both protein tyrosine phosphorylation and histamine release in a dose-dependent manner. These results were consistent with previous studies in rodent mast cells. In contrast, no tyrosine phosphorylation of phospholipase C gamma1 and Btk (Bruton's tyrosine kinase) were observed in our experimental conditions. These results suggest that the greater part of the early and late signaling events in HCMC is similar to those obtained with rodent mast cells and indicated that the requirement of tyrosine phosphorylation in the activation process of each of the signaling molecules might be different in HCMC and rodent mast cells. Our finding indicates that HCMC may be useful for analysis of Fc epsilonRI-mediated signal transduction in human mast cells.
Suzuki
H
H
Pharmaceutical Research Laboratory, Kirin Brewery Co., Ltd., Gunma, Japan. su-hidefumi@kirin.co.jp
Takei
M
M
Yanagida
M
M
Nakahata
T
T
Kawakami
T
T
Fukamachi
H
H
D048868:Adaptor Proteins, Signal Transducing; D001798:Blood Proteins / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D004789:Enzyme Activation / Q000276:immunology; D004792:Enzyme Precursors / Q000378:metabolism; D005312:Fetal Blood; D019833:Genistein / Q000494:pharmacology; D006636:Histamine Release; D006801:Humans; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes / Q000378:metabolism; D008407:Mast Cells / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism; D051966:Phospholipase C gamma; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D050721:Proto-Oncogene Proteins c-cbl; D017455:Receptors, IgE / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction / Q000276:immunology*; D013347:Subcellular Fractions / Q000378:metabolism; D000072377:Syk Kinase; D010738:Type C Phospholipases / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases; D019061:src-Family Kinases / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D001798:Blood Proteins; D003432:Cross-Linking Reagents; D004792:Enzyme Precursors; C061909:HCLS1 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes; D011518:Proto-Oncogene Proteins; D017455:Receptors, IgE; D019000:Phosphotyrosine; D019833:Genistein; D050721:Proto-Oncogene Proteins c-cbl; D044767:Ubiquitin-Protein Ligases; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D010738:Type C Phospholipases; D051966:Phospholipase C gamma; C067419:CBL protein, human; D002118:Calcium