IgE-anti-IgE-induced prostaglandin D2 release from cultured human mast cells.
To elucidate the characterization of cultured human mast cells, the generation of prostaglandin D2 (PGD2) upon stimulation with IgE-anti IgE challenge was determined by using gas chromatography/mass spectrometry/selected ion monitoring (GC/MS/SIM) assay. Mononuclear cells obtained from human umbilical cord blood were cultured in the presence of Steel Factor, IL-6, and PGE2. After 8 weeks of culture, approximately 90 to 95% of cultured cells became tryptase-positive and included basophilic granules. At 12 weeks of culture, cells were harvested and incubated with 1 microgram/ml of human IgE for 1 hr and then challenged with 10 micrograms/ml of anti-IgE. The level of PGD2 release into the supernatant was measured by GC/MS/SIM with the stable isotope dilution method. Cyclooxygenase inhibitors affected the PGD2 release from the cell by IgE-anti IgE challenge incubation. Indomethacin inhibited 98% of PGD2 release. Acetylsalicylic acid (aspirin) and NS-398 (potent COX-2 inhibitor) also inhibited PGD2 release by 85 and 45% respectively.
Obata
T
T
Division of Molecular Cell Biology, Jikei University School of Medicine, Tokyo, Japan.
Nagakura
T
T
Kanbe
M
M
Masaki
T
T
Maekawa
K
K
Yamashita
K
K
D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D053818:Chymases; D005312:Fetal Blood / Q000166:cytology; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007073:Immunoglobulin E / Q000494:pharmacology*; D007150:Immunohistochemistry; D008407:Mast Cells / Q000276:immunology* / Q000378:metabolism* / Q000648:ultrastructure; D015230:Prostaglandin D2 / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism; D053802:Tryptases
D000888:Antibodies, Anti-Idiotypic; D007073:Immunoglobulin E; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases; D015230:Prostaglandin D2