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dc.contributor.authorScheibe, Evgenia
dc.contributor.authorLienenklaus, Stefan
dc.contributor.authorMay, Tobias
dc.contributor.authorMagalhães, Vladimir Gonçalves
dc.contributor.authorWeiss, Siegfried
dc.contributor.authorBrinkmann, Melanie M
dc.date.accessioned2013-10-24T14:30:40Z
dc.date.available2013-10-24T14:30:40Z
dc.date.issued2013
dc.identifier.citationMeasurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells. 2013, 1064:355-66 Methods Mol. Biol.en
dc.identifier.issn1940-6029
dc.identifier.pmid23996270
dc.identifier.doi10.1007/978-1-62703-601-6_25
dc.identifier.urihttp://hdl.handle.net/10033/304603
dc.description.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.
dc.language.isoenen
dc.rightsArchived with thanks to Methods in molecular biology (Clifton, N.J.)en
dc.titleMeasurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for Infection Research, Braunschweig, Germany.en
dc.identifier.journalMethods in molecular biology (Clifton, N.J.)en
refterms.dateFOA2014-10-16T00:00:00Z
html.description.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.


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