Main description:

This volume presents standard approaches and the most recent technical advances used to study innate immune activation. Chapters detail the assessment of macrophage activation, measuring innate immune responses to bacterial viability, quantification of secreted proteins, reporter systems, protocols examining specific innate immune activation by TLRs, RLRs, cGAS, and inflammasomes. 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.

Authoritative and practical, Innate Immune Activation: Methods and Protocols aims to be a useful and informative reference tool for further study into this vital field.

Contents:

1 Emerging Concepts in Innate Immunity

Karin Pelka and Dominic De Nardo

2 Bioinformatic Assessment of Macrophage Activation by the Innate Immune System

Thomas Ulas, Joachim L. Schultze, and Marc Beyer

3 Generation of Genetic Knockouts in Myeloid Cell Lines using a Lentiviral CRISPR/Cas9 System

Paul J. Baker and Seth L. Masters

4 Modeling Primary Human Monocytes with the Trans-differentiation Cell Line BLaER1

Moritz M. Gaidt, Francesca Rapino, Thomas Graf, and Veit Hornung

5 Measurement of NF- B Activation in TLR-activated Macrophages

Orna Ernst, Sharat J. Vayttaden, and Iain D.C. Fraser

6 Biochemical Isolation of the Myddosome from Murine Macrophages

Yunhao Tan and Jonathan C. Kagan

7 Generation of Innate Immune Reporter Cell Lines using Retroviral Transduction

Yamel Cardona Gloria and Dominic De Nardo

8 Myddosome Formation by Luminescence-based Mammalian Interactome Mapping (LUMIER)

Olaf-Oliver Wolz, Manfred Koegl, and Alexander N. R. Weber

9 Inflammatory Caspases: Activation and Cleavage of Gasdermin-D in vitro and during Pyroptosis

Yue Zhao, Jianjin Shi and Feng Shao

10 Methods for Detecting ASC and ASC Speck Formation

Florian Hoss, Verena Rolfes, Mariana R Davanso, Tarcio T Braga, and Bernardo S Franklin

11 Measuring Innate Immune Responses to Bacterial Viability

Julien Moretti, Nicolas Vabret, and J. Magarian Blander

12 Methods to Study Cell Swelling-induced Inflammasome Activation

Vincent Compan and Pablo Pelegrin

13 Detecting Release of Bacterial dsDNA into the Host Cytosol using Fluorescence Microscopy

Roland Felix Dreier, Jose Carlos Santos, and Petr Broz

14 Quantitative Proteomics of Secreted Proteins

Annika Frauenstein and Felix Meissner

15 Simultaneous Detection of Cellular Viability and Interleukin-1 Secretion from Single Cells by ELISpot

Stephanie A. Conos, Lisa M. Lindqvist, and James E. Vince

16 Detection and Quantification of MAV Aggregation via Confocal Imaging

Tan A Nguyen, Lachlan Whitehead, and Ken C Pang