Purification of human basophils. Their response to anti-IgE. 1989

M A Lett-Brown, and L Robinson, and H S Juneja, and J A Grant
Department of Internal Medicine, University of Texas Medical Branch, Galveston 77550.

Although usually the least prevalent blood leukocyte, the basophil can release potent soluble factors in response to multiple triggers. We purified basophils from normal volunteers by means of isopycnic centrifugation and affinity binding of mononuclear cells. The majority of the basophils from most subjects were recovered in a band formed between Percoll layers with densities of 1.070 and 1.080; at this stage basophils represented a mean of 22% of total leukocytes. These cells were reacted with monoclonal antibodies to T (OKT-11) and B (anti-HLA-DR) lymphocytes; B and T cells were removed by adsorption to insoluble antibodies against mouse immunoglobulin resulting in a mean purity of 75% basophils with a yield of 54%. These highly enriched basophils resembled unpurified basophils in terms of (1) intracellular histamine content, (2) spontaneous release of histamine in buffer, and (3) percentage of histamine released by anti-IgE. These findings suggest that the techniques used to purify the basophils do not affect the functional integrity of human basophils.

UI MeSH Term Description Entries
D007073 Immunoglobulin E An immunoglobulin associated with MAST CELLS. Overexpression has been associated with allergic hypersensitivity (HYPERSENSITIVITY, IMMEDIATE). IgE
D002469 Cell Separation Techniques for separating distinct populations of cells. Cell Isolation,Cell Segregation,Isolation, Cell,Cell Isolations,Cell Segregations,Cell Separations,Isolations, Cell,Segregation, Cell,Segregations, Cell,Separation, Cell,Separations, Cell
D002499 Centrifugation, Density Gradient Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Centrifugations, Density Gradient,Density Gradient Centrifugation,Density Gradient Centrifugations,Gradient Centrifugation, Density,Gradient Centrifugations, Density
D006632 Histamine An amine derived by enzymatic decarboxylation of HISTIDINE. It is a powerful stimulant of gastric secretion, a constrictor of bronchial smooth muscle, a vasodilator, and also a centrally acting neurotransmitter. Ceplene,Histamine Dihydrochloride,Histamine Hydrochloride,Peremin
D006636 Histamine Release The secretion of histamine from mast cell and basophil granules by exocytosis. This can be initiated by a number of factors, all of which involve binding of IgE, cross-linked by antigen, to the mast cell or basophil's Fc receptors. Once released, histamine binds to a number of different target cell receptors and exerts a wide variety of effects. Histamine Liberation,Histamine Liberations,Histamine Releases
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001491 Basophils Granular leukocytes characterized by a relatively pale-staining, lobate nucleus and cytoplasm containing coarse dark-staining granules of variable size and stainable by basic dyes. Basophil

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