Abstract
The production of cytokines is a crucial element of the host response to viral and bacterial infections. To follow these events in vivo, transgenic mice have become a valuable tool to study cytokine production through induction of reporter genes. We describe here the generation and immortalization of cells derived from transgenic reporter mice for development of a high-throughput assay system for virus- or bacteria-induced cytokine induction. As an example we describe mouse cytomegalovirus (MCMV) infection of immortalized fibroblasts derived from mice expressing the firefly luciferase reporter downstream of the IFN-β promoter. Common methods to determine IFN-β production, including ELISA, quantitative real-time PCR (qPCR), and transient reporter assays using plasmid-based reporter constructs, have disadvantages and limitations. Transient transfections influence type I IFN responses in most cell types, and IFN-β ELISA as well as qPCR are both laborious and expensive. The method presented here is highly sensitive as well as cost-effective, and allows monitoring of a robust and dose-dependent induction of IFN-β upon virus infection in cell lysates as well as living cells.
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References
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Acknowledgements
This work was supported by the Helmholtz Association through grants VH-NG-637 and the Virtual Institute VISTRIE VH-VI-424 issued to M.M.B. We thank Kendra Ann Bussey for critical reading of the manuscript.
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Scheibe, E., Lienenklaus, S., May, T., Magalhães, V.G., Weiss, S., Brinkmann, M.M. (2013). Measurement of Mouse Cytomegalovirus-Induced Interferon-β with Immortalized Luciferase Reporter Cells. In: Bailer, S., Lieber, D. (eds) Virus-Host Interactions. Methods in Molecular Biology, vol 1064. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-601-6_25
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DOI: https://doi.org/10.1007/978-1-62703-601-6_25
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-600-9
Online ISBN: 978-1-62703-601-6
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