Macrophages and Dendritic Cells

1. Front Matter

   Pages i-xii

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2. Generation of Retroviral Macrophage cDNA Expression Libraries and Functional Screening for Surface Receptors

   • Georgia Schäfer, Gordon D. Brown*

   Pages 1-15

3. 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, Paola Ricciardi-Castagnoli

   Pages 17-27

4. Macrophage Activation: Classical Vs. Alternative

   • Andrea Classen, Jorge Lloberas, Antonio Celada

   Pages 29-43

5. Analysis of Macrophage Phagocytosis: Quantitative Assays of Phagosome Formation and Maturation Using High-Throughput Fluorescence Microscopy

   • Benjamin E. Steinberg, Sergio Grinstein

   Pages 45-56

6. Detection of Activated Rab7 GTPase with an Immobilized RILP Probe

   • Jim Sun, Ala-Eddine Deghmane, Cecilia Bucci, Zakaria Hmama

   Pages 57-69

7. Probing In Vivo Origins of Mononuclear Phagocytes by Conditional Ablation and Reconstitution

   • Chen Varol, Limor Landsman, Steffen Jung

   Pages 71-87

8. 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, Siamon Gordon

   Pages 89-101

9. Exploiting Fc Chimaeric Proteins for the Identification of Ligands Specific for the Mannose Receptor

   • Luisa Martinez-Pomares

   Pages 103-122

10. The Expression of Exogenous Genes in Macrophages: Obstacles and Opportunities

    • Xia Zhang, Justin P. Edwards, David M. Mosser

    Pages 123-143

11. Genetic Modification of Murine Dendritic Cells by RNA Transfection

    • Maurizio Ceppi, Enrico Schmidt, Philippe Pierre

    Pages 145-156

12. Recording Phagosome Maturation Through the Real-Time, Spectrofluorometric Measurement of Hydrolytic Activities

    • Robin M. Yates, Albin Hermetter, David G. Russell

    Pages 157-171

13. Derivation and Characterization of Murine Alternatively Activated (M2) Macrophages

    • Victor W.H. Ho, Laura M. Sly

    Pages 173-185

14. Isolation and Culture of Human Hematopoietic Progenitors for Studies of Dendritic Cell Biology

    • Mattias Svensson

    Pages 187-202

15. Generation, Culture and Flow-Cytometric Characterization of Primary Mouse Macrophages

    • Ulrike Schleicher, Christian Bogdan

    Pages 203-224

16. Analyzing Classical and Alternative Macrophage Activation in Macrophage/Neutrophil-Specific IL-4 Receptor-Alpha-Deficient Mice

    • Frank Brombacher, Berenice Arendse, Reagon Peterson, Alexandra Hölscher, Christoph Hölscher

    Pages 225-252

17. Analysis of Matrix Metalloproteinase Secretion by Macrophages

    • Paul T. Elkington, Justin A. Green, Jon S. Friedland

    Pages 253-265

18. 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, Teunis B.H. Geijtenbeek

    Pages 267-285

19. Stable Lentiviral Vector-Mediated Gene Silencing in Human Monocytic Cell Lines

    • Jimmy S. Lee, Neil E. Reiner

    Pages 287-300

20. Use of Fluorescent Probes to Detect Lipid Signaling Intermediates in Macrophages

    • Vivian Kwok, Eric Vachon, Gregory P. Downey

    Pages 301-328

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.

• Acquired immune system
• Activation
• Cell isolation
• Immune System
• Immunity
• Innate immune system
• Macrophages
• Phagocytosis
• Phagosome
• Vivo
• cell biology
• regulation
• Cell culture

From the reviews: “This book, part of the Methods in Molecular Biology series, provides detailed descriptions of techniques that … may be adapted to individual research projects. … a book on state-of-the-art approaches to investigate the roles of macrophages and dendritic cells in health and disease to better understand their function in the host response. … useful for graduate students, postdoctoral fellows, and experienced investigators. … the protocols are described in great detail in this book, which should be helpful to anyone considering establishing a new research procedure.” (Marion C. Cohen, Doody’s Review Service, December, 2009)

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