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A Sensitive and Versatile Cytokine Bioassay Based on Type I Interferon Signaling in 2fTGH Cells

  • Lennart Zabeau
  • José Van der Heyden
  • Jan TavernierEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 820)

Abstract

We have designed a sensitive and versatile bioassay for quantification of series of cytokines. The assay makes use of chimeric receptors composed of the extracellular, ligand-binding part of the cognate cytokine receptor and the transmembrane and cytosolic part of the type I interferon receptor. Receptors can be homo- (e.g. erythropoietin), di- (e.g. interleukin-5), or even trimeric (e.g. interleukin-2). Stable expression of these chimeras in the 2fTGH cell line allows an interferon-type signaling, which makes a positive selection in conditioned medium possible or a negative selection using a toxic guanine analog. The cytokine of interest is quantified by the extent of cell survival or cell toxicity respectively, which can be measured by easy and cheap crystal violet staining. This bioassay is sensitive in the lower picogram per milliliter range and, in contrast to ELISA methods, only measures the concentration of biologically active cytokines. Using this approach, hypersensitive 2fTGH cell lines have been developed for type I and II interferons, erythropoietin, interleukin-2, and interleukin-5.

Key words

Bioassay Chimeric receptors Interferon 2fTGH 6-Thioguanine Toxicity 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Lennart Zabeau
    • 1
  • José Van der Heyden
    • 1
  • Jan Tavernier
    • 1
    Email author
  1. 1.Department of Medical Protein Research, Flanders Institute for BiotechnologyGhent University, Faculty of Medicine and Health SciencesGhentBelgium

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