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(t) 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.
1. Generation of Retroviral Macrophage cDNA Expression Libraries and Functional Screening for Surface Receptors Georgia Schafer and Gordon D. Brown 2. Generation of Murine Growth Factor-Dependent Long-Term Dendritic Cell Lines to Investigate Host-Parasite Interactions Alessandra Mortellaro, Matteo Urbano, Stefania Citterio, Maria Foti, Francesca Granucci, and Paola Ricciardi-Castagnoli 3. Macrophage Activation: Classical Versus Alternative Andrea Classen, Jorge Lloberas, and Antonio Celada 4. Analysis of Macrophage Phagocytosis: Quantitative Assays of Phagosome Formation and Maturation Using High-Throughput Fluorescence Microscopy Benjamin E. Steinberg and Sergio Grinstein 5. Detection of Activated Rab7 GTPase with an Immobilized RILP Probe Jim Sun, Ala-Eddine Deghmane, Cecilia Bucci, and Zakaria Hmama 6. Probing In Vivo Origins of Mononuclear Phagocytes by Conditional Ablation and Reconstitution Chen Varol, Limor Landsman, and Steffen Jung 7. Multivalent Protein Probes for the Identification and Characterization of Cognate Cellular Ligands for Myeloid Cell Surface Receptors Hsi-Hsien Lin, Gin-Wen Chang, Yi-Shu Huang, Cheng-Chih Hsiao, Martin Stacey, and Siamon Gordon 8. Exploiting Fc Chimaeric Proteins for the Identification of Ligands Specific for the Mannose Receptor Luisa Martinez-Pomares 9. The Expression of Exogenous Genes in Macrophages: Obstacles and Opportunities Xia Zhang, Justin P. Edwards, and David M. Mosser 10. Genetic Modification of Murine Dendritic Cells by RNA Transfection Maurizio Ceppi, Enrico Schmidt, and Philippe Pierre 11. Recording Phagosome Maturation Through the Real-Time, Spectrofluorometric Measurement of Hydrolytic Activities Robin M. Yates, Albin Hermetter, and David G. Russell 12. Derivation and Characterization of Murine Alternatively Activated (M2) Macrophages Victor W.H. Ho and Laura M. Sly 13. Isolation and Culture of Human Hematopoietic Progenitors for Studies of Dendritic Cell Biology Mattias Svensson 14. Generation, Culture and Flow-Cytometric Characterization of Primary Mouse Macrophages Ulrike Schleicher and Christian Bogdan 15. Analyzing Classical and Alternative Macrophage Activation in Macrophage/Neutrophil-Specific IL-4 Receptor-Alpha-Deficient Mice Frank Brombacher, Berenice Arendse, Reagon Peterson, Alexandra Holscher, and Christoph Holscher 16. Analysis of Matrix Metalloproteinase Secretion by Macrophages Paul T. Elkington, Justin A. Green, and Jon S. Friedland 17. Identification of Pathogen Receptors on Dendritic Cells to Understand their Function and to Identify New Drug Targets Lot de Witte, Marein A.W.P. de Jong, Jeroen den Dunnen, Yvette van Kooyk, and Teunis B.H. Geijtenbeek 18. Stable Lentiviral Vector-Mediated Gene Silencing in Human Monocytic Cell Lines Jimmy S. Lee and Neil E. Reiner 19. Use of Fluorescent Probes to Detect Lipid Signaling Intermediates in Macrophages Vivian Kwok, Eric Vachon, and Gregory P. Downey 20. Large Scale Phagosome Preparation Adrien F. Vinet and Albert Descoteaux 21. Coordinate Regulation of Sphingosine Kinase and Actin Dynamics Shankar S. Iyer and David J. Kusner