Chapter title |
cGAMP Quantification in Virus-Infected Human Monocyte-Derived Cells by HPLC-Coupled Tandem Mass Spectrometry
|
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Chapter number | 9 |
Book title |
Innate Antiviral Immunity
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7237-1_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7236-4, 978-1-4939-7237-1
|
Authors |
Jennifer Paijo, Volkhard Kaever, Ulrich Kalinke, Paijo, Jennifer, Kaever, Volkhard, Kalinke, Ulrich |
Abstract |
Upon virus infection, cells of the innate immune system such as dendritic cells and macrophages can mount type I interferon (IFN-I) responses that restrict viral dissemination. To inform host cells of virus infection, detection of cytosolic DNA is one important mechanism. Inappropriate sensing of endogenous DNA and subsequent induction of IFN-I responses can also cause autoimmunity, highlighting the need to tightly regulate DNA sensing. The cyclic GMP-AMP synthase (cGAS) was recently identified to be the major sensor of cytosolic DNA that triggers IFN-I expression. Upon DNA binding, cGAS synthesizes the second messenger cyclic guanosine-adenosine monophosphate (cGAMP) that induces IFN-I expression by the activation of the stimulator of interferon genes (STING). Notably, cGAMP does not only act in infected cells, but can also be relocated to noninfected bystander cells to there trigger IFN-I expression. Thus, direct quantification of cGAMP in cells of the innate immune system is an important approach to study where, when, and how DNA is sensed and IFN-I responses are induced. Here, we describe a method that allows specific quantification of cGAMP from extracts of virus-infected human myeloid cells by HPLC-coupled tandem mass spectrometry. |
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