title: Eosinophils maintain their capacity to signal and release eosinophil cationic protein upon repetitive stimulation with the same agonist.
abstract: Eosinophils contain in their granules eosinophil cationic protein (ECP) and other basic proteins that have been implicated in immunity to parasites and pathophysiology of chronic allergic responses. In a model of eosinophil degranulation, we show that eosinophils release ECP upon short-term GM-CSF priming and stimulation with either platelet-activating factor (PAF) or the anaphylatoxin C5a, but not eotaxin. Restimulation with the same agonist (PAF or C5a) was unsuccessful as assessed by monitoring intracellular calcium concentration and ECP release. In contrast, upon an intermediate washing step, eosinophils rapidly transduced PAF and C5a signals followed by significant ECP releases. Ligand-binding studies demonstrated that only a proportion of PAF receptors is internalized upon cell stimulation and that washing of the cells removes the agonist from the cell surface. Upon repetitive stimulation, eosinophils with less than 50% of the original ECP content were obtained. Such eosinophils did not increase cellular ECP levels even in the presence of the eosinophil survival factor GM-CSF in overnight cultures. In vivo studies revealed that eosinophils always express detectable amounts of ECP under chronic inflammatory conditions. In conclusion, we have shown that eosinophils maintain their capacity to degranulate upon repetitive stimulation with the same agonist as long as the receptor is not occupied from a previous stimulation. The cellular content of ECP appears to be a no limiting factor in the case of repetitive stimulation, implying that mature eosinophils may not require a significant ECP resynthesis.
authors.affiliation: Swiss Institute of Allergy and Asthma Research, University of Zurich, Davos, Switzerland.
authors.forename: H U
authors.initials: HU
authors.lastname: Simon
authors.forename: M
authors.initials: M
authors.lastname: Weber
authors.forename: E
authors.initials: E
authors.lastname: Becker
authors.forename: Y
authors.initials: Y
authors.lastname: Zilberman
authors.forename: K
authors.initials: K
authors.lastname: Blaser
authors.forename: F
authors.initials: F
authors.lastname: Levi-Schaffer
mesh_terms: D001798:Blood Proteins / Q000819:agonists* / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D054413:Chemokine CCL11; D019742:Chemokines, CC; D015936:Complement C5a / Q000502:physiology; D016207:Cytokines / Q000502:physiology; D003600:Cytosol / Q000378:metabolism; D003888:Desensitization, Immunologic; D047091:Eosinophil Granule Proteins; D004802:Eosinophilia / Q000097:blood / Q000378:metabolism / Q000473:pathology; D004804:Eosinophils / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D009928:Organ Specificity / Q000235:genetics; D010972:Platelet Activating Factor / Q000502:physiology; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D012260:Ribonucleases; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology*; D013997:Time Factors
chemical_list: D001798:Blood Proteins; C515456:CCL11 protein, human; D054413:Chemokine CCL11; D019742:Chemokines, CC; D016207:Cytokines; D047091:Eosinophil Granule Proteins; D010972:Platelet Activating Factor; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C039351:platelet activating factor receptor; D015936:Complement C5a; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D012260:Ribonucleases; D002118:Calcium