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Updated: Aug 8, 2026
Basophil Activation Test for Investigation of IgE-Mediated Mechanisms in Drug Hypersensitivity
Published on: September 16, 2011
Flow cytometric evaluation of human basophils
1Institut National de Transfusion Sanguine, Hôpital Rothschild, Paris, France.
Insights
Researchers identified human basophils using flow cytometry by detecting low CD45 and high IgE cell surface antigens. This method allowed for effective isolation and characterization of these crucial immune cells.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Leukocyte surface antigen expression is critical for immune cell identification.
- Basophils play a key role in allergic responses and immune modulation.
- Accurate identification of basophils is essential for immunological research.
Purpose of the Study:
- To characterize human basophils based on CD45 and IgE cell surface antigen expression.
- To develop and validate a method for isolating basophils using flow cytometry.
- To investigate the influence of divalent cations on basophil antigen expression.
Main Methods:
- Flow cytometry was employed to analyze CD45 and IgE expression on human leukocytes.
- Immunomagnetic depletion using anti-IgE antibodies and streptavidin-coated magnetic beads was performed.
- Morphological analysis confirmed the identity of isolated cell populations.
Main Results:
- A distinct subset of leukocytes expressing low CD45 (CD45dim) and high IgE (IgEbright) was identified as basophils (>80%).
- Immunomagnetic depletion effectively isolated this CD45dim-IgEbright subset (>90% depletion).
- Presence of calcium and magnesium significantly altered CD45 and IgE expression, decreasing IgE and increasing CD45.
Conclusions:
- CD45dim-IgEbright phenotype is a reliable marker for human basophil identification via flow cytometry.
- Immunomagnetic depletion provides an efficient method for basophil isolation.
- Divalent cations modulate CD45 and IgE expression on basophils, impacting their surface antigen profile.
Abstract:
The expression of CD45 and IgE cell surface antigens on human leucocytes was studied by flow cytometry. More than 80% of sorted cells that expressed low CD45 (CD45dim) and high IgE (IgEbright) antigen site density were identified as basophils. Immunomagnetic depletion of the CD45dim-IgEbright cell subset by a biotin-coupled anti-IgE antibody and streptavidin-coated magnetic beads was greater than 90%, and more than 80% of cells binding significant numbers of beads exhibited the morphological characteristics of basophils. Interestingly, when the cell staining was performed in the presence of calcium and magnesium, we observed a significant increase of CD45 and an equivalent decrease of IgE cell surface expression, as well as an IgE concentration dependent diminution of the number of CD45dim-IgEbright cells.
