Innate antiviral immunity : methods and protocols

✍ Scribed by Mossman, Karen

• Publisher: Humana Press ; Springer
• Year: 2017
• Tongue: English
• Leaves: 277
• Series: Methods in molecular biology 1656
• Category: Library

✦ Synopsis

This book explores methods to study the complex and evolving interplay between a virus and its host that range from model systems to the detection of chemical molecules. The collection starts with the application of humanized mice and zebrafish as model organisms to study virus-host interactions and induction of innate immune responses. Subsequent chapters outline diverse methods to detect small interfering RNAs, microRNAs, and virus-derived dsRNA from a variety of cells, tissues, and organisms, as well as to interrogating the cytosolic RNA and DNA sensing pathways, including using RNA PAMPs as molecular tools, purification of cGAMP from virus particles and infected cells, and mechanisms to visualize the subcellular localization and activation of the adaptor proteins MAVS and STING. Cutting-edge methods, including high-throughput and genome-wide CRISPR/Cas9 screens, chromosome conformation capture, and whole-exome sequencing, are described to identify novel mediators, pathways, and variants underlying host susceptibility. Given the importance of studying these pathways and players under physiologic conditions, methods describing the isolation of primary mouse sensory neurons and group 2 innate lymphoid cells are also provided. Finally, this collection comes full circle back to the whole organism level and concludes with epidemiological methods to investigate virus-host interactions and the induction of innate immunity. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Innate Antiviral Immunity: Methods and Protocols spans a diverse array of approaches to study and elucidate the intricacies of this vital area of study. Read more...

Abstract:

This book explores methods to study the complex and evolving interplay between a virus and its host that range from model systems to the detection of chemical molecules. Read more...

✦ Table of Contents

Front Matter ....Pages i-xi
The Application of Humanized Mouse Models for the Study of Human Exclusive Viruses (Fatemeh Vahedi, Elizabeth C. Giles, Ali A. Ashkar)....Pages 1-56
Zebrafish as a Model for the Study of Host-Virus Interactions (Peng Fei Zou, Pin Nie)....Pages 57-78
Northern Blot Detection of Virus-Derived Small Interfering RNAs in Caenorhabditis elegans Using Nonradioactive Oligo Probes (Tianyun Long, Rui Lu)....Pages 79-88
Extraction and qPCR-Based Detection of miRNAs from Cultured PBMCs of Bubaline Origin (Chandra S. Mukhopadhyay, Ramneek Verma, Jasdeep Singh)....Pages 89-102
Visualizing Virus-Derived dsRNA Using Antibody-Independent and -Dependent Methods (Sarah J. Poynter, Stephanie J. DeWitte-Orr)....Pages 103-118
RNA PAMPs as Molecular Tools for Evaluating RIG-I Function in Innate Immunity (Renee C. Ireton, Courtney Wilkins, Michael Gale Jr.)....Pages 119-129
Methods to Visualize MAVS Subcellular Localization (Christine Vazquez, Dia C. Beachboard, Stacy M. Horner)....Pages 131-142
Purification of Cyclic GMP-AMP from Viruses and Measurement of Its Activity in Cell Culture (Alice Mayer, Jonathan Maelfait, Anne Bridgeman, Jan Rehwinkel)....Pages 143-151
cGAMP Quantification in Virus-Infected Human Monocyte-Derived Cells by HPLC-Coupled Tandem Mass Spectrometry (Jennifer Paijo, Volkhard Kaever, Ulrich Kalinke)....Pages 153-166
Methods of Assessing STING Activation and Trafficking (Vladislav Pokatayev, Nan Yan)....Pages 167-174
Genome-Wide CRISPR/Cas9 Screening for High-Throughput Functional Genomics in Human Cells (Shiyou Zhu, Yuexin Zhou, Wensheng Wei)....Pages 175-181
High-Throughput Screening for Identification of Novel Innate Immune Activators (Bryan J. Gall, Victor R. DeFilippis)....Pages 183-193
Chromosome Conformation Capture for Research on Innate Antiviral Immunity (Yoon Jung Kim, Tae Hoon Kim)....Pages 195-208
Discovery of Variants Underlying Host Susceptibility to Virus Infection Using Whole-Exome Sequencing (Gabriel A. Leiva-Torres, Nestor Nebesio, Silvia M. Vidal)....Pages 209-227
Isolation, Purification, and Culture of Primary Murine Sensory Neurons (Sarah Katzenell, Jorge R. Cabrera, Brian J. North, David A. Leib)....Pages 229-251
Isolation of Group 2 Innate Lymphoid Cells from Mouse Lungs (Claudia U. Duerr, Jörg H. Fritz)....Pages 253-261
Epidemiological Methods (Biao Wang, Mark Loeb)....Pages 263-271
Back Matter ....Pages 273-275

✦ Subjects

Natural immunity -- Laboratory manuals;Natural immunity;Immunity, Innate;Virus Diseases -- immunology