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A sensitive and versatile bioassay for ligands that signal through receptor clustering

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Abstract
The induced expression of xanthine-guanine phosphoribosyl transferase (XGPRT) by low concentrations (similar to 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 similar to 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.
Keywords
INTERFERON-ALPHA, ALPHA-BETA RECEPTOR, CELL-LINE, GM-CSF, EXPRESSION, ERYTHROPOIETIN, CLONING, ASSAY, INTERLEUKIN-5, ESTABLISHMENT

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Chicago
Van Ostade, Xaveer, Liesbeth Schauvliege, Els Pattyn, Annick Verhee, Joël Vandekerckhove, and Jan Tavernier. 2000. “A Sensitive and Versatile Bioassay for Ligands That Signal Through Receptor Clustering.” Journal of Interferon and Cytokine Research 20 (1): 79–87.
APA
Van Ostade, X., Schauvliege, L., Pattyn, E., Verhee, A., Vandekerckhove, J., & Tavernier, J. (2000). A sensitive and versatile bioassay for ligands that signal through receptor clustering. JOURNAL OF INTERFERON AND CYTOKINE RESEARCH, 20(1), 79–87.
Vancouver
1.
Van Ostade X, Schauvliege L, Pattyn E, Verhee A, Vandekerckhove J, Tavernier J. A sensitive and versatile bioassay for ligands that signal through receptor clustering. JOURNAL OF INTERFERON AND CYTOKINE RESEARCH. 2000;20(1):79–87.
MLA
Van Ostade, Xaveer, Liesbeth Schauvliege, Els Pattyn, et al. “A Sensitive and Versatile Bioassay for Ligands That Signal Through Receptor Clustering.” JOURNAL OF INTERFERON AND CYTOKINE RESEARCH 20.1 (2000): 79–87. Print.
@article{127727,
  abstract     = {The induced expression of xanthine-guanine phosphoribosyl transferase (XGPRT) by low concentrations (similar to 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 similar to 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.},
  author       = {Van Ostade, Xaveer and Schauvliege, Liesbeth and Pattyn, Els and Verhee, Annick and Vandekerckhove, Jo{\"e}l and Tavernier, Jan},
  issn         = {1079-9907},
  journal      = {JOURNAL OF INTERFERON AND CYTOKINE RESEARCH},
  keyword      = {INTERFERON-ALPHA,ALPHA-BETA RECEPTOR,CELL-LINE,GM-CSF,EXPRESSION,ERYTHROPOIETIN,CLONING,ASSAY,INTERLEUKIN-5,ESTABLISHMENT},
  language     = {eng},
  number       = {1},
  pages        = {79--87},
  title        = {A sensitive and versatile bioassay for ligands that signal through receptor clustering},
  url          = {http://dx.doi.org/10.1089/107999000312757},
  volume       = {20},
  year         = {2000},
}

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