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Flow cytometry for basophil activation markers: The measurement of CD203c up-regulation is as reliable as CD63 expression in the diagnosis of cat allergy
- Source :
- Journal of Immunological Methods. 320:40-48
- Publication Year :
- 2007
- Publisher :
- Elsevier BV, 2007.
-
Abstract
- The flow cytometric basophil activation test (BAT), based on the detection of allergen-induced CD63 expression, has been proved effective in the diagnosis of various IgE-mediated allergies. However, there is not yet consensus about the suitability of CD203c expression as a specific basophil activation marker and its diagnostic reliability. The goal of the present study was to compare measurement of CD63 and CD203c expression using BAT in a model of cat allergy and to determine optimal experimental conditions for both markers. Heparinized whole blood samples from 20 cat allergic patients and 19 controls were incubated with Fel d1 (relevant allergen) or anti-FcepsilonRI (positive control) either in IL-3 or IL-3-free conditions. An optimal gating of basophils was achieved in triple staining protocols: anti-IgE PE/anti-CD45 PerCP/anti-CD63 FITC or anti-IgE FITC/anti-CD45 PerCP/anti-CD203c PE. We demonstrated that IL-3 significantly enhanced CD63-induced expression by basophils obtained from cat allergic patients in response to Fel d1. Sensitivity was found to be 100%. The CD203c protocol, when performed under IL-3-free conditions, also demonstrated 100% sensitivity. Only one of the control subjects was positive in both tests. In conclusion, using well-defined experimental conditions, the measurement of CD203c up-regulation on basophils in response to specific allergens is as reliable as CD63-BAT for the in vitro diagnosis of patients with IgE-mediated allergy.
- Subjects :
- Leukotrienes
Allergy
Immunology
Basophil Degranulation Test
Enzyme-Linked Immunosorbent Assay
Platelet Membrane Glycoproteins
Basophil
medicine.disease_cause
Flow cytometry
Allergen
Antigen
Antigens, CD
Hypersensitivity
medicine
Animals
Humans
Immunology and Allergy
Pyrophosphatases
Betula
Whole blood
medicine.diagnostic_test
CD63
Phosphoric Diester Hydrolases
Tetraspanin 30
business.industry
Flow Cytometry
medicine.disease
Basophils
Up-Regulation
Basophil activation
medicine.anatomical_structure
Case-Control Studies
Cats
Interleukin-3
business
Biomarkers
Subjects
Details
- ISSN :
- 00221759
- Volume :
- 320
- Database :
- OpenAIRE
- Journal :
- Journal of Immunological Methods
- Accession number :
- edsair.doi.dedup.....a410e1fbb4405d0acb65830c43ed5f13
- Full Text :
- https://doi.org/10.1016/j.jim.2006.12.002