Detection of high affinity receptor sites for IL 1. beta. on a human B lymphoblastoid line which fail to recognize IL 1. cap alpha

Conference · Wed Mar 05 04:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6851399

Chin, J; Cameron, P; Sigal, N H; Schmidt, J A

A large number of EBV-transformed human B lines were screened for their ability to bind human pI 6.8 IL 1 (IL 1..beta..) which was labeled to high specific radioactivity with Bolton-Hunter reagent. One of these, designated 2C2, bound (/sup 125/)I-IL 1 in a saturable dose-dependent fashion. Scatchard analysis of direct binding data obtained at equilibrium suggested a single family of receptor sites, at approx. 10,000 sites per cell, with a K/sub d/ = 1.5 +/- 0.2 (+SD) nM. Competition experiments with cold pI 6.8 IL 1 gave a K/sub i/ = 1.0 +/- 0.3 nM. No competition was seen with a 20-fold molar excess of human IL 2, human gamma-INF, or the pI 5.2 and pI 5.4 species of human IL 1. These anionic species of IL 1 have recently been purified to homogeneity by us from monocyte culture supernatants. Amino acid sequence analysis of the pI 5.4 species demonstrates that it is encoded by the recently reported IL 1..cap alpha.. cDNA. Cross linking of pI 6.8 (/sup 125/)I-IL 1 to intact 2C2 cells with increasing amounts of cross linker revealed a single band with a MW congruent to 80,000. Cross-linking was totally abolished by excess unlabeled pI 6.8 IL 1 but not by excess pI 5.4 IL 1. These results show that the receptor for IL 1..beta.. on 2C2 cells is highly specific for one species of human IL 1 and raises the possibility that IL 1..cap alpha.. and IL 1..beta.., though very similar in their biological properties, have separate receptor sites.

• Research Organization: Merck Research Labs., Rahway, NJ
• OSTI ID: 6851399
• Report Number(s): CONF-8604222-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:4
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CHEMICAL REACTIONS
CONNECTIVE TISSUE CELLS
CPB
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
DOSE-RESPONSE RELATIONSHIPS
ELECTRON CAPTURE RADIOISOTOPES
GROWTH FACTORS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LEUKOCYTES
LYMPHOCYTES
LYMPHOKINES
MAMMALS
MAN
MATERIALS
MEMBRANE PROTEINS
MITOGENS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
POLYMERIZATION
PRIMATES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
SOMATIC CELLS
TRACER TECHNIQUES
VERTEBRATES