THE IDENTIFICATION AND CHARACTERIZATION OF UMBILICAL-CORD BLOOD-DERIVED HUMAN BASOPHILS

Citation
Cl. Kepley et al., THE IDENTIFICATION AND CHARACTERIZATION OF UMBILICAL-CORD BLOOD-DERIVED HUMAN BASOPHILS, Journal of leukocyte biology, 64(4), 1998, pp. 474-483
Citations number
50
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Immunology,"Cell Biology",Hematology
ISSN journal
0741-5400
Volume
64
Issue
4
Year of publication
1998
Pages
474 - 483
Database
ISI
SICI code
0741-5400(1998)64:4<474:TIACOU>2.0.ZU;2-T
Abstract
Cross-linking allergen-specific immunoglobin E on human peripheral blo od basophils results in the release of histamine and other inflammator y mediators that initiate allergy and asthma. The signaling pathways l eading from IgE binding to mediator release have not been well establi shed, mainly due to the difficulty in obtaining adequate numbers of hi ghly purified basophils, It was the goal of this study to easily obtai n Fc epsilon RI-positive human basoplrils in high yield and purity for studies of signal transduction pathways, We describe all in vitro cul ture system in which pulsing normal human cord blood leukocytes with i nterleukin-3 (IL-3) for 3-4 h followed by incubation in medium with fe tal bovine serum generates a cell population that is predominately Fc epsilon RI positive between 14 and 28 days of culture. These cells res emble peripheral blood basophils when examined by light and electron m icroscopy. Like normal blood basophils, they express the integrins, CD 11b, CD18, CD29, and CD49d. A majority of the IL-3-pulsed cells also e xpress a marker recognized by the basophil-specific antibody, 2D7, Fc epsilon RI cross-linking results in a time and dose-dependent release of histamine. Fc epsilon RI cross-linking also stimulates protein-tyro sine phosphorylation, thought to be the first event leading to the IgE -mediated activation of peripheral blood basophils, These studies esta blish cord blood as an accessible source from which basophil-like cell s call be developed to examine Fc epsilon RI-mediated signal transduct ion.