Dendritic Cell Protocols

This updated book presents protocols from experts in the field that cover the basics and more complex forays into the exploration of DC development and function, both in mice and humans. It aims to become a bench-side handbook for both beginners and experts.

Given the vital importance of immune system research, the gathering of clear, consistent, and informative protocols involving the study of dendritic cells is paramount. Bringing the popular first edition fully up to date, Dendritic Cell Protocols, Second Edition presents protocols from experts in the field that cover the basics and more complex forays into the exploration of DC development and function, both in mice and humans. The first section of the volume involving humans explores topics such as the isolation of blood DC subtypes, primary skin Langerhans cells, and the generation of gene-manipulated human DCs with the inclusion of more clinically relevant methods as well, while the second section involving rodent models delves into DC and precursor generation in vitro, isolation ex vivo, disease models, as well as DC functions and properties. Written in the highly successful Methods in Molecular Biology ™ series style, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Dendritic Cell Protocols, Second Edition aims to become a bench-side handbook for both beginners and experts in the field of DC research and a long-term reference for some of the most popular methods put forward by those who lead the field.

Provides step-by-step procedures for DC isolation from rodent and human tissue, as well as easy in vitro generation of equivalent cell types Explores diverse assays to measure DC function in the context of tolerance, autoimmune disease, allergy, and immunity to viral and bacterial infection All assays used currently by leading experts in the field for high impact studies Preluded by a digestible introduction to DC function and development, covering both humans and rodents Includes techniques that are cutting edge and state-of-the-art in the field of DC biology Contains a comprehensive list of models to test the effect that your favourite gene or immunological model has on DC development or function Helps readers to generate a particular specialized DC subtype in vitro saving time with greater focus on a biologically relevant system Examines isolating DC from almost all organs of the mouse Aids in the identification of the myriad subtypes in human and mice for further study Provides a standardized model for the use of human DCs in clinical trials Includes supplementary material: sn.pub/extras

Inhalt
I: Reviews.- Review of Human DC Subtypes.- Review of Murine Dendritic Cells: Types, Location, and Development.- II: Human DC Protocols.- Isolation of Human Blood DC Subtypes.- Isolation of Immature Primary Langerhans Cells from Human Epidermal Skin.- Isolation and In Vitro Generation of Gene-Manipulated Human Plasmacytoid and Conventional Dendritic Cells.- In Vivo Modulation of Gene Expression by Lentiviral Transduction in Human Immune System Rag2?/??c ?/? Mice.- Generation of Stable Th1/CTL-, Th2-, and Th17-Inducing Human Dendritic Cells.- Pharmacological Modification of Dendritic Cells to Promote Their Tolerogenicity in Transplantation.- Towards a Standardized Protocol for the Generation of Monocyte-Derived Dendritic Cell Vaccines.- III: Murine Dendritic Cell Methods.- CD8+, CD8?, and Plasmacytoid Dendritic Cell Generation In Vitro Using flt3 Ligand.- Generation of Large Numbers of Pro-DCs and Pre-DCs In Vitro.- The Generation of Plasmacytoid and Conventional Dendritic Cells with M-CSF.- Isolation of Common Dendritic Cell Progenitors (CDP) from Mouse Bone Marrow.- The Isolation of Mouse Dendritic Cells from Lymphoid Tissues and the Identification of Dendritic Cell Subtypes by Multiparameter Flow Cytometry.- Isolation of Cutaneous Dendritic Cells.- Isolation of Skin Dendritic Cells from Mouse and Man.- Identification and Isolation of Rodent Respiratory Tract Dendritic Cells.- Isolation of Mouse Thymic Dendritic Cell Precursors.- Isolation of Rat Intestinal Lymph DC.- Mouse Models of Viral Infection: Influenza Infection in the Lung.- DCs in Mouse Models of Intracellular Bacterial Infection.- Studying the Function of Dendritic Cells in Mouse Models of Asthma.- Direct Ex Vivo Activation of T cells for Analysis of Dendritic Cells Antigen Presentation.-Methods to Study Pulmonary Dendritic Cell Migration.- Measuring pH, ROS Production, Maturation, and Degradation in Dendritic Cell Phagosomes Using Cytofluorometry-Based Assays.- Adaptive Treg Generation by DCs and Their Functional Analysis.- Generation of Parabiotic Mice for the Study of DC and DC Precursor Circulation.- Defining In Vivo Dendritic Cell Functions Using CD11c-DTR Transgenic Mice.