A new method for studying the binding of human IgE to CD23 and the inhibition of this binding.
CD23, a low-affinity receptor for IgE (Fc epsilonRII), is a type II membrane-bound glycoprotein expressed on many hematopoietic cells, particularly activated B-cells. CD23 binds to IgE at a domain homologous to Ca2+-dependent (C-type) animal lectin. This paper describes a binding assay by which only the specific binding of IgE to CD23 expressed on Epstein-Barr virus (EBV)-transformed B-cell line, L-KT9 cells, can be detected. This assay is useful in the search for CD23 ligands among many chemical compounds, because it is easily carried out and does not require the use of any radiolabeled reagents. Using the assay, we investigated the inhibition of IgE binding to CD23 by fucose-1-phosphate which has been reported to inhibit the binding of sCD23 to IgE [Delespesse, G., Sarfati, M., Wu, C.Y., Fournier, S., Letellier, M., 1992. The low affinity receptor for IgE. Immunol. Rev. 125, 77.] and the binding of CD23 to CD21 [Pochon, S., Graber, P., Yeager, M., Jansen, K., Bernard, A.R., Aubry, T.-P., Bonnefoy, J.-Y., 1992. Demonstration of second ligand for the low affinity receptor for immunoglobulin E (CD23) using recombinant CD23 reconstituted into flourescent liposomes. J. Exp. Med. 176, 389.]. Although both alpha- and beta-L-fucose-l-phosphate/di(cyclohexylammonium) salt decreased the extent of IgE binding to CD23, the inhibitory effects were not due to alpha- or beta-L-fucose-1-phosphate but to cyclohexylamine. The inhibitory effect of cyclohexylamine was dose dependent and the effect was decreased when inhibition tests were carried out in the presence of a 10-fold excess of IgE. These results suggest that cyclohexylamine specifically interacts with the binding of CD23 and IgE.
Sato
T
T
Asaba Branch of Chemicals Research Laboratories, Honen Corporation, Shizuoka, Japan. qzq01553@niftyserve.or.jp
Konishi
A
A
Yasuno
S
S
Arai
J
J
Kamei
M
M
Bitoh
M
M
Yamaguchi
T
T
D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D002461:Cell Line, Transformed; D003514:Cyclohexylamines / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005643:Fucose / Q000031:analogs & derivatives / Q000494:pharmacology; D004854:Herpesvirus 4, Human; D006600:Hexosephosphates / Q000494:pharmacology; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism*; D017455:Receptors, IgE / Q000378:metabolism*
D003514:Cyclohexylamines; D006600:Hexosephosphates; D017455:Receptors, IgE; C081214:fucopyranosyl phosphate; D005643:Fucose; D007073:Immunoglobulin E