The release of basogranulin in response to IgE-dependent and IgE-independent stimuli: validity of basogranulin measurement as an indicator of basophil activation
The release of basogranulin in response to IgE-dependent and IgE-independent stimuli: validity of basogranulin measurement as an indicator of basophil activation
Background: Basogranulin, the novel basophil granule protein recognized by the monoclonal antibody BB1, can be released by stimulation with anti-IgE antibody or calcium ionophore. However, the kinetics and regulation of its secretion are unknown. Objective: We quantified basogranulin and histamine release in response to a range of stimuli to assess whether basogranulin secretion is a reliable marker of basophil activation. Methods: Isolated peripheral blood basophils were stimulated with anti-IgE antibody, calcium ionophore, N -formyl-Met-Leu-Phe, and complement C5a. The released basogranulin and histamine were quantified by dot blotting with BB1 and a fluorometric method, respectively. Basogranulin localization was confirmed by flow cytometry. Results: Both basogranulin and histamine displayed a bell-shaped response curve when basophils were challenged with anti-IgE. Half-maximal release occurred within 30 seconds. Basogranulin levels were maximal by 15 minutes, whereas those for histamine continued increasing to 30 minutes. Wortmannin, a PI3-K inhibitor, suppressed the release of both mediators. Basophils from donors with the “nonreleaser” phenotype secreted neither mediator in response to anti-IgE. Non-IgE-dependent stimuli released both mediators in parallel in a concentration-dependent manner. The correlation between the relative amounts of each mediator released was highly significant (r = .901, P < .0001, N = 87). Flow cytometry revealed that some of the secreted basogranulin adhered to the cell surface. Conclusions: Basogranulin is secreted along with histamine in response to both FcR I-related and unrelated stimuli. It is therefore a valid marker of basophil activation and could provide the basis for an immunoassay that distinguishes between basophil and mast cell activation. (J Allergy Clin Immunol 2003;112:102-8.)
basophil, basogranulin, histamine, ige, a23187, f-met-leu-phe, c5a, wortmannin, nonreleaser, bb1
102-108
Mochizuki, Akinori
44627a24-b50c-4cea-bec8-e290fe19eadb
McEuen, Alan R.
e2054594-c27d-4d45-86ed-c899dc401c3a
Buckley, Mark G.
e4abc0c1-8413-4b27-bded-e94312c66424
Walls, Andrew F.
aaa7e455-0562-4b4c-94f5-ec29c74b1bfe
July 2003
Mochizuki, Akinori
44627a24-b50c-4cea-bec8-e290fe19eadb
McEuen, Alan R.
e2054594-c27d-4d45-86ed-c899dc401c3a
Buckley, Mark G.
e4abc0c1-8413-4b27-bded-e94312c66424
Walls, Andrew F.
aaa7e455-0562-4b4c-94f5-ec29c74b1bfe
Mochizuki, Akinori, McEuen, Alan R., Buckley, Mark G. and Walls, Andrew F.
(2003)
The release of basogranulin in response to IgE-dependent and IgE-independent stimuli: validity of basogranulin measurement as an indicator of basophil activation.
Journal of Allergy and Clinical Immunology, 112 (1), .
(doi:10.1067/mai.2003.1511).
Abstract
Background: Basogranulin, the novel basophil granule protein recognized by the monoclonal antibody BB1, can be released by stimulation with anti-IgE antibody or calcium ionophore. However, the kinetics and regulation of its secretion are unknown. Objective: We quantified basogranulin and histamine release in response to a range of stimuli to assess whether basogranulin secretion is a reliable marker of basophil activation. Methods: Isolated peripheral blood basophils were stimulated with anti-IgE antibody, calcium ionophore, N -formyl-Met-Leu-Phe, and complement C5a. The released basogranulin and histamine were quantified by dot blotting with BB1 and a fluorometric method, respectively. Basogranulin localization was confirmed by flow cytometry. Results: Both basogranulin and histamine displayed a bell-shaped response curve when basophils were challenged with anti-IgE. Half-maximal release occurred within 30 seconds. Basogranulin levels were maximal by 15 minutes, whereas those for histamine continued increasing to 30 minutes. Wortmannin, a PI3-K inhibitor, suppressed the release of both mediators. Basophils from donors with the “nonreleaser” phenotype secreted neither mediator in response to anti-IgE. Non-IgE-dependent stimuli released both mediators in parallel in a concentration-dependent manner. The correlation between the relative amounts of each mediator released was highly significant (r = .901, P < .0001, N = 87). Flow cytometry revealed that some of the secreted basogranulin adhered to the cell surface. Conclusions: Basogranulin is secreted along with histamine in response to both FcR I-related and unrelated stimuli. It is therefore a valid marker of basophil activation and could provide the basis for an immunoassay that distinguishes between basophil and mast cell activation. (J Allergy Clin Immunol 2003;112:102-8.)
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Published date: July 2003
Keywords:
basophil, basogranulin, histamine, ige, a23187, f-met-leu-phe, c5a, wortmannin, nonreleaser, bb1
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Local EPrints ID: 27273
URI: http://eprints.soton.ac.uk/id/eprint/27273
ISSN: 0091-6749
PURE UUID: e6ab5030-b53e-41ed-909b-3eb379d0718c
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Date deposited: 27 Apr 2006
Last modified: 16 Mar 2024 02:38
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Author:
Akinori Mochizuki
Author:
Alan R. McEuen
Author:
Mark G. Buckley
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