Tyrosine phosphorylation is required for eosinophil degranulation induced by immobilized immunoglobulins.
Degranulation of eosinophils and release of toxic granule proteins play key roles in allergic diseases such as bronchial asthma. However, the intracellular signaling mechanisms that trigger eosinophil degranulation remain unclear. In this study, we investigated protein tyrosine kinase (PTK) involvement in the degranulation of human blood eosinophils induced by immobilized Ig. Eosinophils stimulated with Sepharose beads coated with secretory IgA (slgA) or IgG showed rapid increases in the tyrosine phosphorylation of intracellular proteins with molecular masses of 50 to 56, 73, 78, 100, and 105 kDa. The Ig-induced phosphorylation response was not affected by pertussis toxin, a known inhibitor of Ig-dependent eosinophil activation. The tyrosine kinase inhibitors genistein and herbimycin A inhibited both the tyrosine phosphorylation and degranulation responses of eosinophils induced by sIgA- or IgG-coated beads. In contrast, eosinophil degranulation induced by PMA was not affected by genistein. Treatment of eosinophils with the protein phosphatase inhibitor pervanadate induced the phosphorylation of a similar set of intracellular proteins as well as cellular degranulation. Pervanadate also stimulated an increase in phosphoinositide hydrolysis, which was consistent with the activation of a phospholipase C-gamma isoform by this stimulus. Genistein pretreatment blocked the Ig-induced phospholipase C activation, providing evidence for PTK involvement in this reaction. These findings indicate that a PTK-dependent signaling pathway plays an important role in triggering the degranulation responses of human eosinophils to immobilized sIgA and IgG.
Kato
M
M
Department of Immunology, Mayo Clinic, Rochester, MN 55905, USA.
Abraham
R T
RT
Kita
H
H
D015550:Cell Degranulation / Q000276:immunology; D004804:Eosinophils / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology*; D019833:Genistein; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D007136:Immunoglobulins / Q000276:immunology*; D007295:Inositol Phosphates / Q000097:blood / Q000378:metabolism; D007529:Isoflavones / Q000494:pharmacology; D037342:Pertussis Toxin; D010716:Phosphatidylinositols / Q000494:pharmacology; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000097:blood / Q000502:physiology*; D015398:Signal Transduction / Q000276:immunology; D014638:Vanadates / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D007136:Immunoglobulins; D007295:Inositol Phosphates; D007529:Isoflavones; D010716:Phosphatidylinositols; D010566:Virulence Factors, Bordetella; C076715:pervanadate; D007073:Immunoglobulin E; D014638:Vanadates; D019833:Genistein; D037342:Pertussis Toxin; D011505:Protein-Tyrosine Kinases