Measuring inflammasome activation in response to bacterial infection.

Details

Serval ID
serval:BIB_2040569D7217
Type
A part of a book
Publication sub-type
Chapter: chapter ou part
Collection
Publications
Title
Measuring inflammasome activation in response to bacterial infection.
Title of the book
Methods in Molecular Biology
Author(s)
Broz P., Monack D.M.
Publisher
Springer
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Publication state
Published
Issued date
2013
Peer-reviewed
Oui
Volume
1040
Chapter
6
Pages
65-84
Language
english
Abstract
Inflammasomes are multi-protein signaling platforms assembled in response to viral and bacterial pathogens as well as endogenous danger signals. Inflammasomes serve as activation platforms for the mammalian cysteine protease caspase-1, a central mediator of innate immunity. The hallmarks of inflammasome activation are the processing of caspase-1, the maturation and release of interleukin-1β (IL-1β) and the induction of pyroptosis, a lytic inflammatory cell death. This protocol describes methods for studying inflammasome activation in response to bacterial pathogens in bone-marrow derived murine macrophages (BMDMs). In particular, we outline the protocols to measure cytokine maturation by ELISA and pyroptosis by the release of Lactate Dehydrogenase (LDH). In addition, we describe methods to visualize endogenous ASC specks or foci in infected cells and to study the release of processed caspase-1, caspase-11 and mature cytokines into the cell supernatant by Western blotting. General considerations are discussed to design and optimize the infection protocol for the study of inflammasome activation by other bacterial pathogens.
Keywords
Animals, Apoptosis Regulatory Proteins, Bacterial Infections/immunology, Bacterial Infections/metabolism, Blotting, Western, CARD Signaling Adaptor Proteins, Caspase 1/metabolism, Cell Differentiation, Cytokines/metabolism, Cytoskeletal Proteins/metabolism, Inflammasomes/metabolism, Macrophages/cytology, Macrophages/immunology, Macrophages/metabolism, Macrophages/microbiology, Mice, Microscopy, Confocal
Pubmed
Create date
25/10/2017 10:05
Last modification date
29/07/2024 8:59
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