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Innate Immune Activation

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Cover of 'Innate Immune Activation'

Table of Contents

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    Book Overview
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    Chapter 1 Emerging Concepts in Innate Immunity
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    Chapter 2 Bioinformatic Assessment of Macrophage Activation by the Innate Immune System
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    Chapter 3 Generation of Genetic Knockouts in Myeloid Cell Lines Using a Lentiviral CRISPR/Cas9 System
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    Chapter 4 Modeling Primary Human Monocytes with the Trans–Differentiation Cell Line BLaER1
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    Chapter 5 Measurement of NF-κB Activation in TLR-Activated Macrophages
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    Chapter 6 Biochemical Isolation of the Myddosome from Murine Macrophages
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    Chapter 7 Generation of Innate Immune Reporter Cells Using Retroviral Transduction
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    Chapter 8 Examining Myddosome Formation by Luminescence-Based Mammalian Interactome Mapping (LUMIER)
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    Chapter 9 Inflammatory Caspases: Activation and Cleavage of Gasdermin-D In Vitro and During Pyroptosis
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    Chapter 10 Detection of ASC Speck Formation by Flow Cytometry and Chemical Cross-linking
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    Chapter 11 Measuring Innate Immune Responses to Bacterial Viability
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    Chapter 12 Methods to Study Cell Swelling-Induced Inflammasome Activation
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    Chapter 13 Detecting Release of Bacterial dsDNA into the Host Cytosol Using Fluorescence Microscopy
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    Chapter 14 Quantitative Proteomics of Secreted Proteins
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    Chapter 15 Simultaneous Detection of Cellular Viability and Interleukin-1β Secretion from Single Cells by ELISpot
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    Chapter 16 Detection and Quantification of MAVS Aggregation via Confocal Microscopy
Attention for Chapter 11: Measuring Innate Immune Responses to Bacterial Viability
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Chapter title
Measuring Innate Immune Responses to Bacterial Viability
Chapter number 11
Book title
Innate Immune Activation
Published in
Methods in molecular biology, January 2018
DOI 10.1007/978-1-4939-7519-8_11
Pubmed ID
Book ISBNs
978-1-4939-7518-1, 978-1-4939-7519-8
Authors

Julien Moretti, Nicolas Vabret, J. Magarian Blander

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.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 8 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 8 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 3 38%
Professor 1 13%
Student > Bachelor 1 13%
Student > Master 1 13%
Unknown 2 25%
Readers by discipline Count As %
Agricultural and Biological Sciences 3 38%
Immunology and Microbiology 2 25%
Unknown 3 38%