Measurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells.

2.50
Hdl Handle:
http://hdl.handle.net/10033/304603
Title:
Measurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells.
Authors:
Scheibe, Evgenia; Lienenklaus, Stefan; May, Tobias; Magalhães, Vladimir Gonçalves; Weiss, Siegfried; Brinkmann, Melanie M
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.
Affiliation:
Helmholtz Centre for Infection Research, Braunschweig, Germany.
Citation:
Measurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells. 2013, 1064:355-66 Methods Mol. Biol.
Journal:
Methods in molecular biology (Clifton, N.J.)
Issue Date:
2013
URI:
http://hdl.handle.net/10033/304603
DOI:
10.1007/978-1-62703-601-6_25
PubMed ID:
23996270
Type:
Article
Language:
en
ISSN:
1940-6029
Appears in Collections:
publications of the research group viral immunomodulation (VIMM)

Full metadata record

DC FieldValue Language
dc.contributor.authorScheibe, Evgeniaen
dc.contributor.authorLienenklaus, Stefanen
dc.contributor.authorMay, Tobiasen
dc.contributor.authorMagalhães, Vladimir Gonçalvesen
dc.contributor.authorWeiss, Siegfrieden
dc.contributor.authorBrinkmann, Melanie Men
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.en
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

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