The basophil activation test by flow cytometry: recent developments in clinical studies, standardization and emerging...

Area of Science:

• Immunology
• Allergy Diagnostics

Background:

• Immediate allergy diagnosis relies on clinical history, skin tests, and specific IgE.
• Functional in vitro tests are needed for complex allergy cases, mimicking allergen-basophil interaction.
• Histamine release tests have limitations in sensitivity and specificity for basophil functional responses.

Purpose of the Study:

• To review advancements in basophil activation testing (BAT) using flow cytometry.
• To highlight the role of anti-CRTH2/DP2 antibodies in basophil identification.
• To present a standardized triple staining protocol for improved BAT.

Main Methods:

• Utilizing flow cytometry to detect basophil activation markers (CD63, CD203c) upon allergen challenge.
• Employing a novel triple staining protocol for enhanced basophil recognition.
• Investigating the use of anti-CRTH2/DP2 antibodies for specific basophil identification.

Main Results:

• Flow cytometry offers a reliable method for monitoring basophil activation.
• The new triple staining protocol standardizes the basophil activation test.
• The improved BAT is suitable for both allergy diagnosis and pharmacological research.

Conclusions:

• Flow cytometry-based basophil activation testing is a standardized tool for in vitro immediate allergy diagnosis.
• This method is also valuable for pharmacological studies on human basophils.
• Further multicenter studies are needed to clinically assess BAT in large populations and establish cut-off values.