Publication
Title
A sensitive and versatile bioassay for ligands that signal through receptor clustering
Author
Abstract
The induced expression of xanthine-guanine phosphoribosyl transferase (XGPRT) by low concentrations (2 pg/ml) of interferon-alpha (IFN-alpha) or IFN-beta in the 2fTGH cell line caused a 50% cytotoxicity when these cells were grown in medium containing 6-thioguanine. We extended the application of this sensitive, reliable, and easy bioassay to other members of the cytokine family. To activate the IFN signaling pathway, we made receptor chimeras, consisting of the IFN type I receptor intracellular and transmembrane domains, fused to either the interleukin-5 (IL-5) receptors or erythropoietin (Epo) receptor extracellular domains as model systems. 2fTGH cells, stably transfected with these receptor chimeras, responded to very low concentrations of IL-5 or Epo (IC50 values of 15 pg and 3 pg/ml, respectively) and thus can be used as a very sensitive bioassay for both ligands. Background activity of IL-5, Epo, tumor necrosis factor (TNF), IL-6, or leptin on cells that did not carry the receptor chimeras was very low. This methodology can in principle be extended to any ligand that acts via clustering of its receptors.
Language
English
Source (journal)
Journal of interferon and cytokine research
Publication
2000
Volume/pages
20:1(2000), p. 79-87
ISI
000085012900009
Full text (Publisher's DOI)
UAntwerpen
Faculty/Department
Research group
Publication type
Subject
External links
Web of Science
Record
Identification
Creation 10.06.2009
Last edited 16.11.2017