The identification and characterization of umbilical cord blood-derived human basophils.
Cross-linking allergen-specific immunoglobin E on human peripheral blood basophils results in the release of histamine and other inflammatory mediators that initiate allergy and asthma. The signaling pathways leading from IgE binding to mediator release have not been well established, mainly due to the difficulty in obtaining adequate numbers of highly purified basophils. It was the goal of this study to easily obtain Fc epsilonRI-positive human basophils in high yield and purity for studies of signal transduction pathways. We describe an in vitro culture system in which pulsing normal human cord blood leukocytes with interleukin-3 (IL-3) for 3-4 h followed by incubation in medium with fetal bovine serum generates a cell population that is predominately Fc epsilonRI positive between 14 and 28 days of culture. These cells resemble peripheral blood basophils when examined by light and electron microscopy. Like normal blood basophils, they express the integrins, CD11b, CD18, CD29, and CD49d. A majority of the IL-3-pulsed cells also express a marker recognized by the basophil-specific antibody, 2D7. Fc epsilonRI cross-linking results in a time and dose-dependent release of histamine. Fc epsilonRI cross-linking also stimulates protein-tyrosine phosphorylation, thought to be the first event leading to the IgE-mediated activation of peripheral blood basophils. These studies establish cord blood as an accessible source from which basophil-like cells can be developed to examine Fc epsilonRI-mediated signal transduction.
Kepley
C L
CL
Department of Pathology, Asthma Research Center, Virginia Commonwealth University/Medical College of Virginia, Richmond, USA. ckepley@rapunzel.unm.edu
Pfeiffer
J R
JR
Schwartz
L B
LB
Wilson
B S
BS
Oliver
J M
JM
D000485:Allergens; D000818:Animals; D015703:Antigens, CD / Q000032:analysis / Q000096:biosynthesis; D001491:Basophils / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D002417:Cattle; D002462:Cell Membrane / Q000276:immunology / Q000502:physiology / Q000648:ultrastructure; D002478:Cells, Cultured; D003470:Culture Media; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000502:physiology; D007231:Infant, Newborn; D016023:Integrins / Q000032:analysis / Q000096:biosynthesis; D007377:Interleukin-3 / Q000494:pharmacology / Q000502:physiology; D008855:Microscopy, Electron, Scanning; D011247:Pregnancy; D017452:Receptors, IgG / Q000032:analysis / Q000096:biosynthesis; D015398:Signal Transduction
D000485:Allergens; D015703:Antigens, CD; D003470:Culture Media; D016023:Integrins; D007377:Interleukin-3; D017452:Receptors, IgG; D007073:Immunoglobulin E