Flow cytometric evaluation of human basophils.
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.
Gane
P
P
Institut National de Transfusion Sanguine, Hôpital Rothschild, Paris, France.
Pecquet
C
C
Lambin
P
P
Abuaf
N
N
Leynadier
F
F
Rouger
P
P
D000888:Antibodies, Anti-Idiotypic; D001426:Bacterial Proteins; D001491:Basophils / Q000276:immunology*; D001710:Biotin; D002118:Calcium; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis* / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000032:analysis* / Q000276:immunology; D008274:Magnesium; D008863:Microspheres; D019809:Streptavidin
D000888:Antibodies, Anti-Idiotypic; D001426:Bacterial Proteins; D007073:Immunoglobulin E; D001710:Biotin; D019809:Streptavidin; D017493:Leukocyte Common Antigens; D008274:Magnesium; D002118:Calcium