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Basophil Activation Test for Investigation of IgE-Mediated Mechanisms in Drug Hypersensitivity
Published on: September 16, 2011
[Analysis of membrane expression of the CD63 human basophil activation marker. Applications to allergologic
J Sainte-Laudy1, C Vallon, J C Guérin
1Département des maladies respiratoires, Hôpital de la Croix-Rousse, Lyon.
Insights
A new flow cytometry method analyzes human basophil activation using anti-IgE and anti-CD63. This technique shows promise for diagnosing hypersensitivity reactions, particularly hapten hypersensitivity.
Area of Science:
- Immunology
- Cellular Biology
- Allergy Diagnostics
Background:
- Basophil activation is a key indicator in allergic responses.
- Accurate methods for assessing basophil activation are crucial for diagnosing hypersensitivity.
- Current diagnostic methods for certain hypersensitivities, like hapten hypersensitivity, require improvement.
Purpose of the Study:
- To develop and validate a novel flow cytometry method for analyzing human basophil activation.
- To assess the utility of this method in pharmacological studies and hypersensitivity diagnosis.
- To evaluate CD63 expression as a marker for basophil activation.
Main Methods:
- A flow cytometry technique was established using anti-IgE and anti-CD63 double labeling.
- Human basophils were stimulated with anti-IgE to assess activation.
- CD63 expression was analyzed on the membrane of activated basophils.
- Results were compared with histamine release measurements.
Main Results:
- The flow cytometry method demonstrated reliable statistical characteristics for activation percentages.
- Activation percentages obtained were comparable to those from histamine release assays.
- CD63 expression was detected in response to a low-affinity allergen (sulfonyl-HSA conjugate), with healthy donors showing negative results.
- The method is automatable.
Conclusions:
- The developed flow cytometry method is suitable for pharmacological studies of basophil activation.
- This technique shows potential as a reliable diagnostic tool for hapten hypersensitivity.
- CD63 expression serves as a valid marker for basophil activation in this context.
Abstract:
On the basis of the CD 63 bi-modal expression on the membrane of activated basophils, we set up a flow cytometric method for the analysis of human basophils activation by an anti-IgE and anti-CD 63 double labelling. We demonstrated that the statistical characteristics of the percentages of activation obtained by an anti-IgE stimulation allowed the use of this method for pharmacological studies. The percentages of activation were of the same order of magnitude than those obtained by histamine release. CD 63 expression was also observed for a low affinity allergen such as the sulfonyl-HSA conjugate used for sulfites hypersensibility diagnosis, healthy donors being negative. This method, which can be automatized may represent an interesting candidate in the field of hapten hypersensitivity which lacks of reliable diagnostical methods.
