Measuring inflammasome activation in response to bacterial infection.

Détails

ID Serval
serval:BIB_2040569D7217
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Measuring inflammasome activation in response to bacterial infection.
Périodique
Methods in Molecular Biology
Auteur(s)
Broz P., Monack D.M.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Statut éditorial
Publié
Date de publication
2013
Peer-reviewed
Oui
Volume
1040
Pages
65-84
Langue
anglais
Résumé
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.

Mots-clé
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
Création de la notice
25/10/2017 11:05
Dernière modification de la notice
20/08/2019 13:56
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