Changes in filament actin accompanying IgE-dependent and -independent histamine release from IL-3-dependent cultured human basophils.
When cord blood mononuclear cells were cultured in the presence of rhIL-3 for 5 weeks or more, 40-90% of cultured cells became morphologically mature basophils. We analyzed the kinetics of histamine release, changes in filament actin (F-actin), and movement of intracellular Ca2+ ([Ca2+]i) induced by IgE-dependent (anti-IgE) and -independent (fMLP) stimuli in these cultured basophils. Anti-IgE and fMLP released 24.5 +/- 5.4% and 14.5 +/- 4.5% histamine from the cells, respectively. Anti-IgE caused actin polymerization with a peak response at 15 min, which began much later than the elevation of [Ca2+]i. In contrast to anti-IgE stimulation, fMLP induced rapid actin polymerization with a peak response at 30 s in correlation with kinetics of histamine release. Our results indicate that cord blood-derived cultured basophils show similar cell functions to mature basophils, and are useful models with which to investigate the mechanisms of degranulation, specifically when a large amount of highly purified cells are required.
Ebisawa
M
M
Division of Allergy, National Children's Medical Research Center, Tokyo, Japan.
Saito
H
H
Reason
D C
DC
Sakaguchi
N
N
Katsunuma
T
T
Iikura
Y
Y
D000199:Actins / Q000378:metabolism*; D001491:Basophils / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000502:physiology*; D007377:Interleukin-3 / Q000494:pharmacology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology
D000199:Actins; D007377:Interleukin-3; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D009240:N-Formylmethionine Leucyl-Phenylalanine; D002118:Calcium