Title
A sensitive and versatile bioassay for ligands that signal through receptor clustering A sensitive and versatile bioassay for ligands that signal through receptor clustering
Author
Faculty/Department
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences . Biomedical Sciences
Publication type
article
Publication
Subject
Human medicine
Source (journal)
Journal of interferon and cytokine research
Volume/pages
20(2000) :1 , p. 79-87
ISSN
0197-8357
ISI
000085012900009
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
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.
E-info
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