Main description:

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.

Contents:

Preface...
Table of Contents...
Contributing Authors...

1. A Brief Introduction to Effector-Triggered Immunity
Thomas A. Kufer and Maria Kaparakis-Liaskos

2. Lambda Red Recombineering in Shigella flexneri
Adrian Herod, Julie Ryu, and John Rohde

3. Using Genomic Deletion Mutants to Investigate Effector-Triggered Immunity during Legionella pneumophila Infection
Rachelia R. Wibawa, Pengfei Li, Kathleen McCaffrey, and Elizabeth L. Hartland

4. Methods of Bacterial Membrane Vesicle Production, Purification, Quantification, and Examination of their Immunogenic Functions
Natalie J. Bitto and Maria Kaparakis-Liaskos

5. Gene Gun-Mediated Transient Gene Expression for Functional Studies in Plant Immunity
Franz Leissing, Anja Reinstadler, Hannah Thieron, and Ralph Panstruga

6. Host Induced Gene Silencing using BPMV on Soybean to Study Genes in the Soybean Rust Fungus Phakopsora pachyrhizi
Tobias I. Link

7. Virus-Mediated Protein Overexpression (VOX) in Monocots to Identify and Functionally Characterize Fungal Effectors
Kostya Kanyuka

8. Time-Resolved Fluorescence Microscopy Screens on Host Protein Subversion during Bacterial Cell Invasion
Lisa Sanchez, Yuen-Yan Chang, Nora Mellouk, and Jost Enninga

9. Assaying RIPK2 Activation by Complex Formation
Heidrun Steinle, Kornelia Ellwanger, and Thomas A. Kufer

10. Identification of Novel Endogenous NOD Ligands: Quantitative Analysis of Binding Affinities of NOD1 or NOD2 with Sphingosine-1-Phosphate using Microscale Thermophoresis
Gang Pei

11. A Surface Plasmon Resonance-Based Strategy to Characterize Interactions of NLR Proteins with Associated Factors
Jonas Moecking and Matthias Geyer

12. Functional Assessment of Disease-Associated Pyrin Variants
Daria Chirita, Yvan Jamilloux, Thomas Henry, and Flora Magnotti

13. An Assay for the Seeding of Homotypic Pyrin Domain Filament Transitions
Inga V. Hochheiser and Matthias Geyer

14. Detection of Gasdermin Activation and Lytic Cell Death during Pyroptosis and Apoptosis
Benjamin Demarco, Saray Ramos, and Petr Broz

15. Visualizing Effector Triggered Immunity in Response to Pore-Forming Toxins by Live-Cell Imaging
Seong H. Chow and Thomas Naderer

16. Non-Apoptotic Cell Death Control of Neutrophil Extracellular Trap Formation
Akshay A. D'Cruz, Maria Ericsson, and Ben A. Croker

17. Analysis of Bacteria-Triggered Inflammasome-Activation in Neutrophils by Immunoblot
Remi Planes, Karin Santoni, and Etienne Meunier

18. Quantifying Regulated Mitochondrial Fission in Macrophages
Syeda Farhana Afroz, Nicholas D. Condon, Matthew J. Sweet, and Ronan Kapetanovic

19. Live Cell Imaging of T Cell Pyroptosis
Andreas Linder and Veit Hornung

Subject Index List...