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Recognition of the ligand-binding site of human neutrophil (N) receptors for leukotriene B/sub 4/ (LTB/sub 4/) by antibodies to idiotypes (anti-Ids) of mouse monoclonal anti-LTB/sub 4/

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:5454615
; ; ; ;

Monoclonal IgG/sub 2b/ anti-LTB/sub 4/, that binds (/sup 3/H)LTB/sub 4/ with a specificity identical to that of N chemotactic receptors, was purified by affinity chromatography on agarose-protein A and agarose-hexylamide-LTB/sub 4/ for immunization of six rabbits. After rabbits had received 100 ..mu..g of monoclonal anti-LTB/sub 4/ 5 times, the serums of three inhibited the binding of (/sup 3/H)LTB/sub 4/ to anti-LTB/sub 4/ by 50% at dilutions of 1/30-1/480. IgG from serum with anti-Ids inhibited the specific binding of (/sup 3/H)LTB/sub 4/ to Ns by a mean of 22% and 40% (n = 2), at respective concentrations of 0.23 mg/ml and 0.64 mg/ml, whereas normal IgG had no effect. In contrast, anti-Ids and normal IgG inhibited to the same extent the binding of (/sup 3/H)N-f-Met-Leu-Phe to Ns. Analyses of the concentration-dependence of binding of (/sup 3/H)LTB/sub 4/ to Ns pretreated 90 min at 4/sup 0/C with 1.2 mg/ml of anti-Ids, normal IgG, or buffer alone showed that only anti-Ids altered significantly the expression of high-affinity receptors, without substantially affecting the properties of low-affinity receptors. Anti-Ids, but not normal IgG, stimulated N chemotaxis and increases in cytosolic (Ca/sup 2 +/), as quantified by Quin 2 fluorescence, with half-maximal responses at 0.5 mg/ml and 0.005 mg/ml, respectively. The anti-Ids resemble LTB/sub 4/ in binding preferentially to high-affinity receptors and eliciting N responses.

Research Organization:
Univ. of California Medical Center, San Francisco
OSTI ID:
5454615
Report Number(s):
CONF-8604222-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:3; ISSN FEPRA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ALKALINE EARTH METALS
ANIMALS
ANTIBODIES
ARACHIDONIC ACID
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CALCIUM
CARBOXYLIC ACIDS
CARDIOVASCULAR AGENTS
DRUGS
ELEMENTS
IMMUNE REACTIONS
INHIBITION
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LEUKOCYTES
LIGANDS
MAMMALS
MATERIALS
MEMBRANE PROTEINS
METALS
MICE
MONOCARBOXYLIC ACIDS
MONOCLONAL ANTIBODIES
NEUTROPHILS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
PURIFICATION
RABBITS
REACTION KINETICS
RECEPTORS
RODENTS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VASOCONSTRICTORS
VERTEBRATES