New JRI study published in Immunity maps human innate immune cells. To read more, click here. Drs. Julie Magarian Blander and Steven Lipkin are 2018 Daedalus Fund for Innovation recipients. To read more, click here. Donor gift establishes Friedman Center for Nutrition and Inflammation. To read more, click here. Latest Iliev Lab study published online  in Cell Host and MicrobeTo read more, click here. Latest Longman Lab study published in Immunity. To read more, click here. 

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Measuring Innate Immune Responses to Bacterial Viability.

TitleMeasuring Innate Immune Responses to Bacterial Viability.
Publication TypeJournal Article
Year of Publication2018
AuthorsMoretti, J, Vabret, N, J Blander, M
JournalMethods Mol Biol
Volume1714
Pagination167-190
Date Published2018
ISSN1940-6029
Abstract

The innate immune system directly senses microbial viability via the detection of a special class of viability-associated pathogen-associated molecular patterns (vita-PAMPs), such as prokaryotic messenger RNA. In the case of Gram-negative bacteria, detection of bacterial viability by phagocytes leads to a unique activation of inflammasome and type I interferon pathways, resulting in a robust pro-inflammatory innate response and a vigorous adaptive immune response. This protocol describes the methods required to study activation of both inflammasome and type I interferon pathways after stimulation of mouse bone marrow-derived macrophages with live or killed Gram-negative and Gram-positive bacteria. It covers the generation and handling of bone marrow-derived macrophages, the culture and killing of bacteria, the preparation of bacterial messenger RNA, and the stimulation of macrophages with live or killed bacteria. Lastly, this protocol describes the techniques employed to measure the hallmarks of inflammasome (secretion of interleukin-1β) and type I interferon (activation of TBK1, IRF3 and secretion of type I interferon) pathways.

DOI10.1007/978-1-4939-7519-8_11
Alternate JournalMethods Mol. Biol.
PubMed ID29177862