Flow cytometric evaluation of human basophils

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 CD45d'"-IgEbT'ght 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 CD45d'"-IgEbnght cells.

Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-dependent upregulation

“…The most prominent markers are the tetraspanin CD63 [46] and the ecto-enzyme CD203c [47][48][49]. Both markers are suitable to analyze allergen-induced basophil activation and are routinely used in allergy diagnosis [19][20][21][22][23][24][25][26][27][28][29][30]. In the search for additional markers, we analyzed a panel of workshop antibodies for differential reactivity with resting and activated basophils.…”

“…In addition, activation markers like the granuleassociated molecule CD63 and the ecto-enzyme CD203c are upregulated to the plasma membrane in response to FcεRI cross linkage [19][20][21][22][23][24][25][26][27][28][29][30]. These markers are now routinely used for flow cytometry-based basophil activation tests.…”

CCR3 as a single selection marker compared to CD123/HLADR to isolate basophils in flow cytometry: Some comments

“…pos cells are basophils (11). So, operator has to move the gate to capture basophils but, when doing so, he must face these technical issues.…”

Hymenoptera-Venom-Induced Upregulation of the Basophil Activation Marker Ecto-Nucleotide Pyrophosphatase/Phosphodiesterase 3 in Sensitized Individuals

“…Recently, flow cytometry-based tests have been developed that rely on the binding of fluorochrome-conjugated allergen to specific cell-bound IgE or allergen-induced cell surface expression of basophilic granule antigens [4, 5, 6, 7, 8, 9]. Most of the reports describe the induced cell surface expression of the granule-associated molecule CD63 on the cell surface of activated basophils [6, 9].…”