Simplified purification of human basophils.
Studies of human basophils have been limited by the low number present in peripheral blood and the difficulties of purification to homogeneity with reasonable yield and functional status. Reproducible purification of human basophils to 96.5+/-0.5% with a yield of 40.8+/-5.3% was obtained by negative selection using immunomagnetic beads following initial separation by density gradient centrifugation. Isolated cells demonstrated complete viability by vital dye exclusion and spontaneous histamine release following incubation of <5%. Stimulation with anti-IgE or calcium ionophore A23187 caused histamine release and leukotriene C(4) production. Basophils demonstrated dose-dependent chemotaxis to monocyte chemotactic protein-3. This simplified methodology results in fully functional basophils in very high purity and good yield.
Tsang
S
S
UBC Pulmonary Research Laboratory, St. Paul's Hospital, 1081 Burrard Street, Vancouver, B.C., Canada.
Hayashi
M
M
Zheng
X
X
Campbell
A
A
Schellenberg
R R
RR
D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology; D001491:Basophils / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D000001:Calcimycin / Q000494:pharmacology; D002499:Centrifugation, Density Gradient; D054410:Chemokine CCL7; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D016207:Cytokines; D006636:Histamine Release / Q000187:drug effects; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D018189:Immunomagnetic Separation / Q000379:methods*; D066298:In Vitro Techniques; D007476:Ionophores / Q000494:pharmacology; D017997:Leukotriene C4 / Q000096:biosynthesis; D018945:Monocyte Chemoattractant Proteins / Q000494:pharmacology
D000888:Antibodies, Anti-Idiotypic; C515450:CCL7 protein, human; D054410:Chemokine CCL7; D016207:Cytokines; D007476:Ionophores; D018945:Monocyte Chemoattractant Proteins; D017997:Leukotriene C4; D007073:Immunoglobulin E; D000001:Calcimycin