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Flow cytometric evaluation of human basophils
1Institut National de Transfusion Sanguine, Hôpital Rothschild, Paris, France.
Cytometry
|January 1, 1993
Summary
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.