/sup 3/H arachidonic acid incorporation and metabolism in purified human basophils
A central feature of the allergic response is the generation of arachidonic acid (AA) metabolites by basophils and mast cells. In addition, AA metabolism may play a role in regulating the anti-IgE mediated degranulation of human basophils. To study this biochemistry, purified human basophils (>80%) were labeled with /sup 3/H-AA (0.3 ..mu..M, 25 ..mu..Ci/ml, 2 hours at 37/sup 0/C) and subsequently challenged with anti-IgE. Basophils were found to incorporate 45 +/- 3% of the exogenous AA which distributed into phospholipids (PL) (77.1 +/- 3.5%) and neutral lipids (19.7 +/- 3.3%) with only 5.3 +/- 2.7% remaining as the free acid (n = 7). Phosphatidylcholine (23.9 +/- 1.7%), phosphatidylinositol (22.0 +/- 1.4%) and phosphatidylethanolamine (14.5 +/- 2.7%) accounted for the majority of the AA with the remaining PL containing <3%. Anti-IgE (0.1 ..mu..g/ml) challenge led to the release of histamine (23.8 +/- 4.7%) and /sup 3/H-AA (8.1 +/- 1.7%) (n = 5). HPLC analysis revealed unmetabolized /sup 3/H-AA, /sup 3/H-LTC/sub 4/, /sup 3/H-HETE and an unidentified peak which migrated in the prostaglandin region of the elution profile. The same metabolites were released when the basophils were challenged with antigen. The calcium ionophore A23187 (1..mu..g/ml) also caused the release of histamine (37.4 +/- 4.1%) and /sup 3/H-AA (17.0 +/- 2.9%), while the phorbol ester, TPA caused HR (19.7 +/- 5.8%) but no increase in /sup 3/H AA turnover. Because of limited cell numbers this is the first time the authors have been able to study AA metabolism in human basophils.
- Research Organization:
- Johns Hopkins Univ. School of Medicine, Baltimore, MD
- OSTI ID:
- 5325475
- Report Number(s):
- CONF-8604222-; TRN: 86-030488
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ARACHIDONIC ACID
METABOLISM
ALLERGY
ANTIGENS
BASOPHILS
CALCIUM
HISTAMINE
LIQUID COLUMN CHROMATOGRAPHY
MAN
MAST CELLS
METABOLITES
PHORBOL ESTERS
PHOSPHOLIPIDS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ALKALINE EARTH METALS
AMINES
ANIMAL CELLS
ANIMALS
AZOLES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CARCINOGENS
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
ELEMENTS
ESTERS
HETEROCYCLIC COMPOUNDS
IMIDAZOLES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LEUKOCYTES
LIPIDS
MAMMALS
MATERIALS
METALS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PATHOLOGICAL CHANGES
PRIMATES
SEPARATION PROCESSES
SOMATIC CELLS
VERTEBRATES
550501* - Metabolism- Tracer Techniques