Measurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells.
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Magalhães, Vladimir Gonçalves
Brinkmann, Melanie M
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AbstractThe 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.
CitationMeasurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells. 2013, 1064:355-66 Methods Mol. Biol.
AffiliationHelmholtz Centre for Infection Research, Braunschweig, Germany.
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