Differential release of mediators from human basophils: Differences in arachidonic acid metabolism following activation by unrelated stimuli
We have examined the release of histamine and LTC4 from purified human basophils challenged with several different stimuli, both physiological and nonphysiological. Basophils (n = 16) challenged with 0.1 micrograms/ml anti-IgE released 38 +/- 4% of their available histamine and 39 +/- 12 ng LTC4/10(6) basophils within 15-30 min. F-Met peptide (n = 8) caused the release of 54 +/- 8% histamine and 42 +/- 25 ng LTC4/10(6) basophils within a period of 2-5 min. C5a caused the release of 22 +/- 3% histamine from selected donors but failed to initiate any LTC4 release unless combined with D2O or 5 mM extracellular calcium. The two nonphysiological stimuli A23187 and TPA caused extensive histamine release, 67 +/- 8 and 82 +/- 11%, respectively, and while A23187 initiated a large and rapid release of leukotriene, TPA failed to release any LTC4 even when combined with D2O or 2-5 mM extracellular calcium. Increased concentrations of extracellular calcium enhanced anti-IgE and f-Met peptide induced release of LTC4 but inhibited the A23187 induced release of leukotriene. A single peak of immunoreactive leukotriene C4 that comigrated with the authentic standard was identified using HPLC followed by radioimmunoassay. No LTD4 or LTE4 could be detected. Purified human basophils incubated with 0.2 microM (3H)AA incorporated 290 pmol/10(6) cells, or 32 +/- 5% of the available label within 60 min. The (3H)AA was taken principally into the phospholipids (73 +/- 5%), with 20 +/- 3% as neutral lipid, and only 5 +/- 2% remaining as the free acid. Three phospholipid subclasses, phosphatidylcholine, PC (24 +/- 2%), phosphatidylinositol, PI (22 +/- 1%), and phosphatidylethanolamine, PE (15 +/- 3%), accounted for the majority of the incorporated (3H)AA while the remainder of the phospholipids accounted for less than 5% of the total cpm.
- Research Organization:
- Johns Hopkins Univ. School of Medicine, Baltimore, MD (USA)
- OSTI ID:
- 5722446
- Journal Information:
- J. Leukocyte Biol.; (United States), Vol. 45:6
- Country of Publication:
- United States
- Language:
- English
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ARACHIDONIC ACID
METABOLISM
HISTAMINE
SECRETION
PHORBOL ESTERS
BIOLOGICAL EFFECTS
BASOPHILS
CALCIUM
COMPLEMENT
DEUTERIUM
IMMUNOGLOBULINS
LIQUID COLUMN CHROMATOGRAPHY
MAN
PEPTIDES
PHENYLALANINE
PHOSPHOLIPIDS
RADIOIMMUNOASSAY
STIMULI
TRITIUM COMPOUNDS
ALKALINE EARTH METALS
AMINES
AMINO ACIDS
ANIMALS
AZOLES
BIOASSAY
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CARCINOGENS
CHROMATOGRAPHY
ELEMENTS
ESTERS
GLOBULINS
HETEROCYCLIC COMPOUNDS
HYDROGEN ISOTOPES
IMIDAZOLES
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LEUKOCYTES
LIGHT NUCLEI
LIPIDS
MAMMALS
MATERIALS
METALS
MONOCARBOXYLIC ACIDS
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PRIMATES
PROTEINS
RADIOASSAY
RADIOIMMUNOLOGY
SEPARATION PROCESSES
STABLE ISOTOPES
TRACER TECHNIQUES
VERTEBRATES
550501* - Metabolism- Tracer Techniques