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

Scheibe, Evgenia
; 
Lienenklaus, Stefan
; 
May, Tobias
; 
Magalhães, Vladimir Gonçalves
; 
Weiss, Siegfried
; 
Brinkmann, Melanie M
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Issue Date
2013
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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.
Citation
Measurement of mouse cytomegalovirus-induced interferon-β with immortalized luciferase reporter cells. 2013, 1064:355-66 Methods Mol. Biol.
Affiliation
Helmholtz Centre for Infection Research, Braunschweig, Germany.
Publisher
Journal
Methods in molecular biology (Clifton, N.J.)
URI
http://hdl.handle.net/10033/304603
DOI
10.1007/978-1-62703-601-6_25
PubMed ID
23996270
PubMed Central ID
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Type
Article
Language
en
Description
Series/Report no.
ISSN
1940-6029
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Archived with thanks to Methods in molecular biology (Clifton, N.J.)
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publications of the junior research group immunomodulation (VIMM)