Macrophages and Dendritic Cells

Expert researchers contribute laboratory protocols involving these two vital cell types at the junction of the innate and acquired immune systems. The volume looks at isolation and cell culturing and then covers topics such as phagocytosis and lipid signaling.

In light of the critical contributions of macrophages and dendritic cells to diverse inflammatory diseases and to immunity and host defense, state-of-the-art approaches to the investigation of their behavior are essential. In Macrophages and Dendritic Cells: Methods and Protocols , expert researchers contribute laboratory protocols involving these two vital cell types functioning at the junction of the innate and acquired immune systems. The volume delves first into isolation and cell culturing then continues with topics such as phagocytosis, genetic manipulation, macrophage activation, and lipid signaling. Written in the highly successful Methods in Molecular Biology ™ series format, chapters include brief introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Macrophages and Dendritic Cells: Methods and Protocols provides a timely and useful guide for both seasoned investigators and neophytes pursuing this imperative field of study.

Provides an easily accessible reference volume for state-of-the-art experimentation by leading researchers in the field Covers a wide range of approaches for the isolation and characterization of macrophages and dendritic cells Provides detailed step-by-step protocols to modify gene expression in cells that are notoriously resistant to genetic manipulation Provides approaches for detailed quantitative functional analyses including activation states and lipid signaling Includes supplementary material: sn.pub/extras

Inhalt
Generation of Retroviral Macrophage cDNA Expression Libraries and Functional Screening for Surface Receptors.- Generation of Murine Growth Factor-Dependent Long-Term Dendritic Cell Lines to Investigate Host-Parasite Interactions.- Macrophage Activation: Classical Vs. Alternative.- Analysis of Macrophage Phagocytosis: Quantitative Assays of Phagosome Formation and Maturation Using High-Throughput Fluorescence Microscopy.- Detection of Activated Rab7 GTPase with an Immobilized RILP Probe.- Probing In Vivo Origins of Mononuclear Phagocytes by Conditional Ablation and Reconstitution.- Multivalent Protein Probes for the Identification and Characterization of Cognate Cellular Ligands for Myeloid Cell Surface Receptors.- Exploiting Fc Chimaeric Proteins for the Identification of Ligands Specific for the Mannose Receptor.- The Expression of Exogenous Genes in Macrophages: Obstacles and Opportunities.- Genetic Modification of Murine Dendritic Cells by RNA Transfection.- Recording Phagosome Maturation Through the Real-Time, Spectrofluorometric Measurement of Hydrolytic Activities.- Derivation and Characterization of Murine Alternatively Activated (M2) Macrophages.- Isolation and Culture of Human Hematopoietic Progenitors for Studies of Dendritic Cell Biology.- Generation, Culture and Flow-Cytometric Characterization of Primary Mouse Macrophages.- Analyzing Classical and Alternative Macrophage Activation in Macrophage/Neutrophil-Specific IL-4 Receptor-Alpha-Deficient Mice.- Analysis of Matrix Metalloproteinase Secretion by Macrophages.- Identification of Pathogen Receptors on Dendritic Cells to Understand their Function and to Identify New Drug Targets.- Stable Lentiviral Vector-Mediated Gene Silencing in Human Monocytic Cell Lines.- Use of Fluorescent Probes to Detect Lipid Signaling Intermediates in Macrophages.- Large-Scale Phagosome Preparation.- Coordinate Regulation of Sphingosine Kinase and Actin Dynamics.