WISH cell line: From the antiviral system to a novel reporter gene assay to test the potency of human IFN-α and IFN-β - Archive ouverte HAL Access content directly
Journal Articles Journal of Immunological Methods Year : 2012

WISH cell line: From the antiviral system to a novel reporter gene assay to test the potency of human IFN-α and IFN-β

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Abstract

Interferons (IFNs) are potent biologically active proteins that are widely used as biopharmaceuticals, so their potency must be correctly identified. Usually, the biological activity is quantified by a bioassay based on its capacity to induce an antiviral state in target cells, but this type of assays is subject to virus manipulation-related issues and they show considerable intra- and inter-assay variability. In this work, we generated a reporter gene assay (RGA) supported on the WISH-Mx/eGFP reporter cell line to determine human type I IFN activity. WISH cells were stably transfected with the enhanced green fluorescent protein (eGFP) gene under the control of type I IFN-inducible Mx2 promoter. This system implies the use of a standardized cell line for human IFN-potency analysis such as WISH cells and the simultaneous use of the sensitive reporter gene eGFP, having also several advantages when compared to antiviral activity assays and other RGAs: it can determine the potency of hIFN-α and hIFN-β using only one cell line showing the highest expression of eGFP after 28h and being only observed in cells treated with type I IFNs due to the specificity of the Mx promoter. It is a sensitive assay and it represents a safe alternative when compared with the conventional antiviral tests. The cell line showed the same sensitivity along 57 generations, allowing its use during two months of successive culture. The inter- and intra-assay coefficients of variation were lower than 20%, demonstrating its reproducibility. In addition, this reporter cell line can be used for the conventional antiviral assay, either for hIFN-α or hIFN-β. In conclusion, we have developed an alternative reporter system for the analysis of type I IFNs, in which its performance make it a suitable candidate to replace or complement conventional bioassays that are currently employed to measure IFN potency.
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Dates and versions

pasteur-00692047 , version 1 (27-04-2012)

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María de Los Milagros Bürgi, Claudio Prieto, Marina Etcheverrigaray, Ricardo Kratje, Marcos Oggero, et al.. WISH cell line: From the antiviral system to a novel reporter gene assay to test the potency of human IFN-α and IFN-β. Journal of Immunological Methods, 2012, epub ahead of print. ⟨10.1016/j.jim.2012.04.010⟩. ⟨pasteur-00692047⟩
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