The purification of human basophils: their immunophenotype and cytochemistry

Abstract

An IgM class monoclonal antibody Bsp-1 that selectively reacts with human basophils was used to label basophils in normal and leukaemic haemopoietic samples. The immunophenotype of Bsp-1+ basophils was determined using a panel of 21 IgG subclass monoclonal antibodies in two-colour immunofluorescence assays. Basophils expressed the leucocyte common antigen, HLA-ABC antigens and antigens defined by CD11 and CD13 monoclonal antibodies. Other myeloid cell (granulocyte-monocyte) associated anti-antigens, lymphoid cell surface determinants and HLA-DR antigens were not detected. Basophil preparations of 95-98% purity were obtained from the peripheral blood of patients with CML and umbilical cords using fluorescence activated cell sorting techniques. Purified basophils exhibited metachromatic staining with toluidine blue, alcain blue and astra blue. PAS staining was observed in 7% of cord Bsp-1+ cells and of 55% CML Bsp-1+ cells. Between 5% and 10% of basophils were chloroacetate esterase-positive which suggests that some Bsp-1+ cells are immature basophils.

Human basophils express CD22 without expression of CD19

“…T-cell activation markers such as CD25, leukocyte adhesion molecules such as CD11b/18, receptors for IgG, BSP-1, IgE, IL-3R␣ (CD123), and several other molecules have been identified on basophils (6,17,(22)(23)(24)(25)(26)(27)(28)(29). But the kinds of antigens expressed on basophils are still a matter of controversy (23,24,27,30). This could be due to paucity of basophils in the peripheral blood.…”

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

“…CD107b was selected because it shares structural similarity and lysosomal expression with CD107a, and is upregulated in activated mast cells in a similar fashion as CD107a [37]. BSP-1 was of interest because it is the first reported cell surface marker that is highly specific for basophils [12,13]. For these experiments, resting and activated basophils were stained with the selected FITC-conjugated antibodies and counter-stained with the CD203c-specific antibody conjugate 97A6-PE.…”

“…This anti-gen was identified as a selective cell surface marker for the basophil and mast cell lineage [10,11]. Apart from CD203c, a number of other basophil-specific markers have been described which are expressed either on the cell surface (Bsp-1) [12,13] or in cytoplasmic compartments (2D7, BB1) [14][15][16] of mature basophils. However, none of these basophil-specific markers are molecularly identified and no information exists about their expression in early basophil and/or mast cell differentiation.…”

Enumeration and immunohistochemical characterisation of bone marrow basophils in myeloproliferative disorders using the basophil specific monoclonal antibody 2D7

“…Basophils express a unique composition of cytoplasmic mediators and cell surface membrane antigens. [10][11][12][13][14][15] Although mast cells and basophils produce several mediators and surface antigens in common, they can clearly be discriminated from each other by their unique profile of granular enzymes and CD molecules. [12][13][14][15][16][17] During the past two decades, the phenotype of human basophils has been analysed extensively using antibodies directed against various leucocyte differentiation antigens.…”