Toll-Like Receptors

by Mccoy, Claire E.; O'Neill, Luke A. J.

• ISBN13: 9781934115725
• ISBN10: 193411572X
• Edition: 1st
• Format: Hardcover
• Copyright: 2009-03-18
• Publisher: Humana Pr Inc

Summary

In addition to their invaluable role in the fight against infection, Toll-Like Receptors (TLRs) in an under- or over-active state can lead to the pathogenesis of disease, making these receptors a key focal point for many research laboratories. In Toll-Like Receptors: Methods and Protocols, experts in the field contribute techniques currently used to study TLRs, their downstream signalling pathways, and their role in the pathogenesis of disease, with sections examining TLR expression and the application of genetic techniques and microarray analysis to TLR research. Written in the highly successful Methods in Molecular Biology'„¢ series format, chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections highlighting tips on troubleshooting and avoiding known pitfalls.Versatile and cutting-edge, Toll-Like Receptors: Methods and Protocols provides an ideal manual for a wide range of biologists and medical researchers studying these vital components of the immune system.

Table of Contents

Preface	p. v
Contributors	p. xi
Methods to Detect and Analyze Toll-Like Receptors
Expression Analysis of the Toll-Like Receptors in Human Peripheral Blood Mononuclear Cells	p. 3
Ligands, Cell-Based Models, and Readouts Required for Toll-Like Receptor Action	p. 15
Toll-Like Receptor Interactions Imaged by FRET Microscopy and GFP Fragment Reconstitution	p. 33
Predicting Toll-Like Receptor Structures and Characterizing Ligand Binding	p. 55
Bioinformatic Analysis of Toll-Like Receptor Sequences and Structures	p. 69
Expression, Purification, and Crystallization of Toll/Interleukin-1 Receptor (TIR) Domains	p. 81
Methods to Analyze Signal Transduction Downstream of Toll-Like Receptor Stimulation
Proteomic Analysis of Protein Complexes in Toll-Like Receptor Biology	p. 91
2D-DIGE: Comparative Proteomics of Cellular Signalling Pathways	p. 105
MAPPIT (Mammalian Protein-Protein Interaction Trap) Analysis of Early Steps in Toll-Like Receptor Signalling	p. 133
Analysis of the Functional Role of Toll-Like Receptor-4 Tyrosine Phosphorylation	p. 145
Analysis of Ubiquitin Degradation and Phosphorylation of Proteins	p. 169
The Generation of Highly Purified Primary Human Neutrophils and Assessment of Apoptosis in Response to Toll-Like Receptor Ligands	p. 191
Cellular Expression of A20 and ABIN-3 in Response to Toll-Like Receptor-4 Stimulation	p. 205
Characterisation of Viral Proteins that Inhibit Toll-Like Receptor Signal Transduction	p. 217
Genetic Techniques in Toll-Like Receptor Analysis
Genetic Dissection of Toll-Like Receptor Signaling Using ENU Mutagenesis	p. 239
Microarray Experiments to Uncover Toll-Like Receptor Function	p. 253
Uncovering Novel Gene Function in Toll-Like Receptor Signalling Using siRNA	p. 277
Genotyping Methods to Analyse Polymorphismsin Toll-Like Receptors and Disease	p. 297
Toll-Like Receptors and Disease
Experimental Models of Acute Infection and Toll-Like Receptor Driven Septic Shock	p. 313
Toll-Like Receptors and Rheumatoid Arthritis	p. 329
Practical Techniques for Detection of Toll-Like Receptor-4 in the Human Intestine	p. 345
Toll-Like Receptor-Dependent Immune Complex Activation of B Cells and Dendritic Cells	p. 363
Innate Immunity, Toll-Like Receptors, and Atherosclerosis: Mouse Models and Methods	p. 381
Generation of Parasite Antigens for Use in Toll-Like Receptor Research	p. 401
Biomarkers Measuring the Activity of Toll-Like Receptor Ligands in Clinical Development Programs	p. 415
Index	p. 441
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