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Proteomic analysis of human eosinophil activation mediated by mast cells, granulocyte macrophage colony stimulating factor and tumor necrosis factor alpha
Author(s) -
LeviSchaffer Francesca,
Temkin Vladislav,
Simon HansUwe,
Kettman JohnR.,
Frey JohannRudolf,
Lefkovits Ivan
Publication year - 2002
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/1615-9861(200211)2:11<1616::aid-prot1616>3.0.co;2-s
Subject(s) - eosinophil , eosinophil peroxidase , tumor necrosis factor alpha , mast cell , major basic protein , granulocyte , granulocyte macrophage colony stimulating factor , microbiology and biotechnology , immunology , biology , interleukin 5 , eosinophil granule proteins , cytokine , chemistry , interleukin , asthma
We assessed mast cell influence on eosinophils, the prominent cells in late and chronic allergic reactions, by comparing the proteomic pattern of eosinophils incubated with mast cells, tumor necrosis factor alpha (TNF‐α) or granulocyte macrophage colony stimulating factor (GM‐CSF). Eosinophils were incubated with the human mast cell line HMC‐1 cellular sonicate and their survival and GM‐CSF production were evaluated. For proteomic studies, eosinophils were cultured with HMC‐1 sonicate, TNF‐α or GM‐CSF in the presence of [ 35 S]methionine, solubilized and submitted to isolelectric focusing separation and sodium dodecyl sulfate polyacrylamide gel electrophoresis in the ISODALT system, followed by radiofluorography and computer image analysis. HMC‐1‐incubated eosinophils displayed increased survival partly mediated by mast cell‐associated TNF‐α, and produced GM‐CSF. Metabolically labeled eosinophils incubated with either HMC‐1, TNF‐α or GM‐CSF released eosinophil peroxidase. Comparison of two‐dimensional gel spots from the eosinophils revealed that each of the three activating signals yielded a distinctly different proteomic pattern of labeled polypeptides. GM‐CSF provided the strongest signal and the highest rate of protein synthesis (1018 spots) followed by TNF‐α (747 spots) and HMC‐1 sonicate (611 spots). A portion of spots differed both in terms of quality and quantity. Although each stimulus induced similar functional effects, the resulting biosynthetic programs of the eosinophils greatly differed. The presented proteomic analysis is the first step in the exploration of molecular mechanisms involved in eosinophil activation.