Innate Antiviral Immunity

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.
The chapter 'Morphological Separation ofClustered Nuclei in Histological Images' is published open access under a CC BY 4.0 license at link.springer.com.

Includes cutting-edge methods and protocols for the study of innate antiviral immunity Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

The Application of Humanized Mouse Models for the Study of Human Exclusive Viruses.- Zebrafish as a Model for the Study of Host-Virus Interactions.- Northern Blot Detection of Virus-Derived Small Interfering RNAs in Caenorhabditis elegans Using Non-Radioactive Oligo Probes.- Extraction and qPCR-Based Detection of miRNAs from Cultured PBMCs of Bubaline Origin.- Visualizing Virus-Derived dsRNA Using Antibody-Independent and -Dependent Methods.- RNA PAMPs as Molecular Tools for Evaluating RIG-I Function in Innate Immunity.- Methods to Visualize MAVS Subcellular Localization.- Purification of Cyclic GMP-AMP from Viruses and Measurement of Its Activity in Cell Culture.- cGAMP Quantification in Virus-Infected Human Monocyte-Derived Cells by HPLC-Coupled Tandem Mass Spectrometry.- Methods of Assessing STING Activation and Trafficking.- Genome-Wide CRISPR/Cas9 Screening for High-Throughput Functional Genomics in Human Cells.- High Throughput Screening for Identification of Novel Innate Immune Activators.- Chromosome Conformation Capture for Research on Innate Antiviral Immunity.- Discovery of Variants Underlying Host Susceptibility to Virus Infection Using Whole-Exome Sequencing.- Isolation, Purification, and Culture of Primary Murine Sensory Neurons.- Isolation of Group 2 Innate Lymphoid Cells from Mouse Lungs.- Epidemiological Methods.