The Inflammasome: Methods and Protocols

ISBN-13: 9781493960118 / Angielski / Miękka / 2016 / 225 str.

Christine M. De Nardo; Eicke Latz

Opis

This Methods in Molecular Biology book offers methods for studying inflammasome function, including generation of inflammasome stimuli, monitoring of caspase-1 activity and processing, activation of IL-1β cytokines, plus lab protocols, material lists and tips.

Spis treści

1. Assessing b-amyloid-induced NLRP3 Inflammasome Activation in Primary Microglia

Mareike Schnaars, Hannes Beckert, and Annett Halle

 

2. Activating the NLRP3 Inflammasome using the Amyloidogenic Peptide IAPP

Clara Westwell-Roper, Aisling Dunne, Man-Lyang Kim, C. Bruce Verchere, and Seth L. Masters

 

3. Assessment and Quantification of Crystal-induced Lysosomal Damage

Peter Duewell and Eicke Latz

 

4. Assessment of Inflammasome Activation in Primary Human Immune Cells

Theo S. Plantinga, Leo A.B. Joosten, and Mihai G. Netea

 

5. NLRP3 Inflammasome Activation and Cytotoxicity Induced by particulate Adjuvants

Marie Yang, Claire H.A. Hearnden, Ewa Oleszycka and Ed C. Lavelle

 

6. Measuring Inflammasome Activation in Response to Bacterial Infection

Petr Broz and Denise M. Monack

 

7. Detection of Pyroptosis by Measuring Released Lactate Dehydrogenase Activity

Manira Rayamajhi, Yue Zhang, and Edward A. Miao

 

8. ASC Speck Formation as a Read-out for Inflammasome Activation

Andrea Stutz, Gabor L. Horvath, Brian G. Monks, and Eicke Latz

 

9. Immunoblotting for Active Caspase-1

Christopher Jakobs, Eva Bartok, Andrej Kubarenko, Franz Bauernfeind and Veit Hornung

 

10. Inflammasome Activation and Inhibition in Primary Murine Bone Marrow-Derived Cells, and Assays for IL-1α, IL-1β and Caspase-1

Katharina S. Schneider¹, Christina J. Thomas, and Olaf Groß

 

11. Reconstituting the NLRP1 Inflammasome in vitro

Benjamin Faustin and John C. Reed

 

12. Assessing ATP Binding and Hydrolysis by NLR Proteins

Jinyao Mo and Joseph A. Duncan

 

13. Blue Native Polyacrylamide Gel Electrophoresis to Monitor Inflammasome Assembly and Composition

Eric M. Kofoed and Russell E. Vance

 

14. Analysis of Microbiota Alterations in Inflammasome-deficient Mice

Eran Elinav, Christoph A Thaiss, and Richard A Flavell

 

15. Quantification of Adipose Tissue Leukocytosis in Obesity

Ryan Grant, Yun-Hee Youm, Anthony Ravussin, and Vishwa Deep Dixit

 

16. In vivo Evaluation of Neutrophil Recruitment in Response to Sterile Particulates

Tamiko Yanagida, Kaoru Orihashi and Hajime Kono

 

Blurb

 Inflammasomes have a seemingly simple architecture. A sensor molecule of the NLR or PYHIN protein family recruits the common adapter molecule apoptosis-associated speck-like protein containing a CARD (ASC). Even so, inflammasome activation is highly regulated at multiple steps and precise details of how inflammasomes are activated at the molecular level remain largely unknown. In The Inflammasome: Methods and Protocols, expert researchers in the field detail methods and protocols to study multiple aspects of inflammasome function. These methods include the generation of inflammasome stimuli, perturbance of cellular homeostasis, assessment of ASC speck formation, monitoring of caspase-1 activity and processing, activation of IL-1β cytokines, and analysis of inflammasome assembly. 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 key tips on troubleshooting and avoiding known pitfalls.