Measurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells.
dc.contributor.author | Scheibe, Evgenia | |
dc.contributor.author | Lienenklaus, Stefan | |
dc.contributor.author | May, Tobias | |
dc.contributor.author | Magalhães, Vladimir Gonçalves | |
dc.contributor.author | Weiss, Siegfried | |
dc.contributor.author | Brinkmann, Melanie M | |
dc.date.accessioned | 2013-10-24T14:30:40Z | |
dc.date.available | 2013-10-24T14:30:40Z | |
dc.date.issued | 2013 | |
dc.identifier.citation | Measurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells. 2013, 1064:355-66 Methods Mol. Biol. | en |
dc.identifier.issn | 1940-6029 | |
dc.identifier.pmid | 23996270 | |
dc.identifier.doi | 10.1007/978-1-62703-601-6_25 | |
dc.identifier.uri | http://hdl.handle.net/10033/304603 | |
dc.description.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. | |
dc.language.iso | en | en |
dc.rights | Archived with thanks to Methods in molecular biology (Clifton, N.J.) | en |
dc.title | Measurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells. | en |
dc.type | Article | en |
dc.contributor.department | Helmholtz Centre for Infection Research, Braunschweig, Germany. | en |
dc.identifier.journal | Methods in molecular biology (Clifton, N.J.) | en |
refterms.dateFOA | 2014-10-16T00:00:00Z | |
html.description.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. |