Toll-Like Receptors (2^nd Ed., 2nd ed. 2016) Practice and Methods Methods in Molecular Biology Series, Vol. 1390

This second edition volume expands on the first edition with new developments on Toll-Like Receptors (TLRs) controlling events such as cross-priming of associated pattern recognition receptors, post-transcriptional regulation, interaction with other cellular and biologic systems, and cancer progression. This book is divided into five sections: Part I outlines methods for TLR detection, interaction, and intracellular trafficking; Part II describes methods and assays to investigate how TLRs cross-prime other pattern recognition receptors, including intracellular DNA receptors and inflammasome formation; Part III highlights RNA regulation, detailing how TLRs can induce RNA transcripts and molecules such as lncRNAs and microRNAs; Part IV explores TLR detection and activation in systems such as epithelial barrier function, metabolism and the circadian clock, as well as cellular systems including T and B lymphocytes; and Part V describes models to delineate the role of TLRs in diseases such as dermatitis, arthritis, and gastric cancer. Written in the highly successful Methods in Molecular Biology series format, each chapter contains a summary, a list of required materials, step-by-step, readily reproducible laboratory protocols, useful notes to investigate TLRs in cell culture, systems and disease, and tips on troubleshooting and avoiding known pitfalls.

Part I: Toll-Like Receptor Detection and Activation

1. Toll-Like Receptors: Ligands, Cell-Based Models, and Readouts for Receptor Action
Jennifer K. Dowling and Jérome Dellacasagrande

2. Bioinformatic Analysis of Toll-Like Receptor Sequences and Structures
Tom P. Monie, Nicholas J. Gay, and Monique Gangloff

3. Toll-Like Receptor Interactions Measured by Microscopic and Flow Cytometric FRET
Gabor L. Horwath, Pia Langhoff, and Eicke Latz

4. Using Confocal Microscopy to Investigate Intracellular Trafficking of Toll-Like Receptors
Harald Husebye and Sarah L. Doyle

5. Assessing the Inhibito

Part II: Toll-Like Receptor Cross-Priming of Associated Receptors

6. Methods for Delivering DNA to Intracellular Receptors
Katryn J. Stacey, Adi Idris, Vitaliya Sagulenko, Nazarii Vitak, and David P. Sester

7. Detection of Interaction between Toll-Like Receptors and Other Transmembrane Proteins by Co-Iummunoprecipitation Assay
Yu-Ran Lee, Won Dae Kang, and You-Me Kim

8. Flow Cytometry-Based Bead-Binding Assay for Measuring Receptor Ligand Specificity
Joris K. Sprokholt, Nina Hertoghs, and Tennis B.H. Geijtenbeek

9. Measuring Monomer-to-Filament

Part III: Toll-Like Receptor Post-Transcriptional Regulation

10. Co-Transcriptomic Analysis by RNA Sequencing to Simultaneously Measure Regulated Gene Expression in Host and Bacterial Pathogen
Timothy Ravasi, Charalampos (Harris) Mavromatis, Nilesh J. Bokil, Mark A. Schembri, and Matthew J. Sweet

11. Simple Methods to Investigate MicroRNA Induction in Response to Toll-Like Receptors
Victoria G. Lyons and Claire E. McCoy

12. Determining the Function of Long Noncoding RNA in Innate Immunity
Susan Carpenter

13. Analysis of Post-Transcriptional Gene Regulation of Nod-Like Receptors via the 3'UTR
Moritz Haneklaus

Part IV: Toll-Like Receptors and System Control

14. TLR Function in Murine CD4+ T Lymphocytes and their Role in Inflammation
Stephanie Flaherty and Joseph M. Reynolds

15. Flow Cytometry Analysis of B Cell Activation and Antibody Responses Induced by Toll-Like Receptors
Egest J. Pone

16. Toll-Like Receptor-Dependent Immune Complex Activation of B Cells and Dendritic Cells
Krishna L. Moody, Melissa B. Uccellini, Ana M. Avalos, Ann Marshak-Rothstein, and Gregory A. Viglianti

17. Analysis of TLR-Induced Metabolic Changes in Dendritic Cells U

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18. Toll-Like Receptor Signaling and the Control of Intestinal Barrier Function
Daniel G.W. Johnston and Sinéad C. Corr

19. Understanding the Role of Cellular Molecular Clocks in Controlling the Innate Immune Response
Anne M. Curtis and Caio T. Fagundes

Part V: Toll-Like Receptors and Disease

20. Methods to Investigate the Role of Toll-Like Receptors in Allergic Contact Dermatitis
Marc Schmidt, Matthias Goebeler, and Stefan F. Martin

21. Allergens and Activation of the Tol

22. Investigating the Role of Toll-Like Receptors in Models of Arthritis
Anna M. Piccinini, Lynn Williams, Fiona E. McCann, and Kim. S. Midwood

23. Delineating the Role of Toll-Like Receptors in the Neuro-Inflammation Model EAE
Franscesca Fallarino, Marco Gargaro, Giada Mondanell, Eric J. Downer, Md Jakir Hossain, and Bruno Gran

24. The Use of MiRNA Antagonists in the Alleviation of Inflammatory Disorders
Lucien P. Garo and Gopal Murugaiyan

25. Investigating the Role of Toll-Like Receptors in Mouse Models of Gastric Cancer
Alison C. West and Brendan J. Jenkins

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

Includes supplementary material: sn.pub/extras