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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 13: Detecting Release of Bacterial dsDNA into the Host Cytosol Using Fluorescence Microscopy
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Chapter title
Detecting Release of Bacterial dsDNA into the Host Cytosol Using Fluorescence Microscopy
Chapter number 13
Book title
Innate Immune Activation
Published in
Methods in molecular biology, January 2018
DOI 10.1007/978-1-4939-7519-8_13
Pubmed ID
Book ISBNs
978-1-4939-7518-1, 978-1-4939-7519-8
Authors

Roland Felix Dreier, José Carlos Santos, Petr Broz

Abstract

Recognition of pathogens by the innate immune system relies on germline-encoded pattern recognition receptors (PRRs) that recognize unique microbial molecules, so-called pathogen-associated molecular patterns (PAMPs). Nucleic acids and their derivatives are one of the most important groups of PAMPs, and are recognized by a number of surface-associated as well as cytosolic PRRs. Cyclic GMP-AMP synthase (cGAS) recognizes the presence of pathogen- or host-derived dsDNA in the cytosol and initiates type-I-IFN production. Here, we describe a methodology that allows for evaluating the association of cGAS with released bacterial dsDNA during Francisella novicida infection of macrophages, by fluorescence confocal microscopy. This method can be adapted to the study of cGAS-dependent responses elicited by other intracellular bacterial pathogens and in other cell types.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 7 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 7 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 4 57%
Student > Bachelor 1 14%
Researcher 1 14%
Unknown 1 14%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 3 43%
Immunology and Microbiology 2 29%
Veterinary Science and Veterinary Medicine 1 14%
Unknown 1 14%