Effector-Triggered Immunity Methods and Protocols

This volume presents a collection of protocols to study effector-triggered immunity (ETI) in both plants and animals from eminent groups in the field. The chapters in this book cover topics such as genetic manipulation of plant and animal pathogens, host cells, and the analysis of key host responses; and techniques used for the analysis of inflammasome activation, cell death pathways, and mitochondria damage in response to pathogens. All of these topics cover a broad spectrum of immunological, biochemical, cell biological, and structural biology approaches to examine ETI. Written in the highly successful Methods in Molecular Biology series format, 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.

Cutting-edge and practical, Effector-Triggered Immunity: Methods and Protocols is a valuable resource for both expert and novice researchers who are interested in learning more about the important and developing field of ETI.

A Brief Introduction to Effector-Triggered Immunity .- Lambda Red Recombineering in Shigella flexneri.- Using Genomic Deletion Mutants to Investigate Effector-Triggered Immunity during Legionella pneumophila Infection.- Methods of Bacterial Membrane Vesicle Production, Purification, Quantification, and Examination of their Immunogenic Functions.- Gene Gun-Mediated Transient Gene Expression for Functional Studies in Plant Immunity.- Host Induced Gene Silencing using BPMV on Soybean to Study Genes in the Soybean Rust Fungus Phakopsora pachyrhizi.- Virus-Mediated Protein Overexpression (VOX) in Monocots to Identify and Functionally Characterize Fungal Effectors.- Time-Resolved Fluorescence Microscopy Screens on Host Protein Subversion during Bacterial Cell Invasion.- Assaying RIPK2 Activation by Complex Formation.- Identification of Novel Endogenous NOD Ligands: Quantitative Analysis of Binding Affinities of NOD1 or NOD2 with Sphingosine-1-Phosphate using Microscale Thermophoresis.- A Surface Plasmon Resonance-Based Strategy to Characterize Interactions of NLR Proteins with Associated Factors.- Functional Assessment of Disease-Associated Pyrin Variants.- An Assay for the Seeding of Homotypic Pyrin Domain Filament Transitions.- Detection of Gasdermin Activation and Lytic Cell Death during Pyroptosis and Apoptosis.- Visualizing Effector Triggered Immunity in Response to Pore-Forming Toxins by Live-Cell Imaging.- Non-Apoptotic Cell Death Control of Neutrophil Extracellular Trap Formation.- Analysis of Bacteria-Triggered Inflammasome-Activation in Neutrophils by Immunoblot.- Quantifying Regulated Mitochondrial Fission in Macrophages.- Live Cell Imaging of T Cell Pyroptosis.