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Leading cell biologists and immunologists present their most useful and innovative techniques for studying the molecular and cellular basis of cell migration in the inflammatory process. The authors provide a series of focused, cutting-edge techniques proceeding from the in vitro analysis of cell migration and the molecular mechanisms underlying this process, to methodologies for the analysis of cell migration in vivo. Experimental approaches useful in establishing the role of cell migration in the pathogenesis of both acute and chronic inflammatory diseases are emphasized. Methods for the analysis of rapid leukocyte adhesion under flow conditions in vitro are described, which may prove especially fruitful for scientists exploring the molecular mechanisms underlying vascular recognition and leukocyte–endothelium interaction. Each fully tested protocol includes an introduction explaining the principle behind the technique, equipment and reagent lists, and tips on troubleshooting and how to avoid known pitfalls.

Ed. D’Ambrosio, Daniele; Sinigaglia, Francesco

cell-migration-in-inflammation-and-immunity-methods-and-protocolscell-migration-in-inflammation-and-immunity-methods-and-protocols.pdf

Table of Contents

1. Chemotaxis and Interaction with Vascular or Lymphatic Endothelium (pp. 1-16)
   Sozzani, Silvano; Vecchi, Annunciata; Allavena, Paola; Mantovani, Alberto
2. Analysis of Integrin-Dependent Rapid Adhesion Under Laminar-Flow Conditions (pp. 17-26)
   Laudanna, Carlo
3. Posttranslational Processing of Chemokines (pp. 27-44)
   Proost, Paul; Mahieu, Frank; Schutyser, Evemie; Van Damme, Jo
4. Chemotactic Profiling of Lymphocyte Subpopulations (pp. 45-52)
   Colantonio, Lucia; Iellem, Andrea; D’Ambrosio, Daniele
5. Measurement of the Levels of Polymerized Actin (F-Actin) in Chemokine-Stimulated Lymphocytes and GFP-Coupled cDNA Transfected Lymphoid Cells by Flow Cytometry (pp. 53-68)
   Vicente-Manzanares, Miguel; Viton, Mariano; Sanchez-Madrid, Francisco
6. Evaluation of Rho Family Small G-Protein Activity Induced by Integrin Ligation on Human Leukocytes (pp. 69-76)
   Gismondi, Angela; Mainiero, Fabrizio; Santoni, Angela
7. Reconstructing Leukocyte Migration in 3D Extracellular Matrix by Time-Lapse Videomicroscopy and Computer-Assisted Tracking (pp. 77-90)
   Friedl, Peter; Brocker, Eva-B.
8. Analyzing Chemotaxis Using Dictyostelium discoideum as a Model System (pp. 91-104)
   Landree, Mark A.; Devreotes, Peter N.
9. Conditional Transgenic Models to Study Chemokine Biology (pp. 105-122)
   Lira, Sergio A.; Mehrad, Borna; Chen, Shu-Cheng; Zalamea, Petronio; Kinsley, David J.; Wiekowski, Maria T.; Coronel, Elizabeth; Vassileva, Galya; Manfra, Denise; Jensen, Kristian K.
10. Intravital Microscopy as a Tool for Studying Recruitment and Chemotaxis (pp. 123-132)
    Cara, Denise C.; Kubes, Paul
11. Tracking Antigen-Specific Lymphocytes In Vivo (pp. 133-146)
    Adams, Claire L.; Rush, Catherine M.; Smith, Karen M.; Garside, Paul
12. Analysis of Homing-Receptor Expression on Infiltrating Leukocytes in Disease States (pp. 147-166)
    Mariani, Margherita; Panina-Bordignon, Paola
13. Interaction of Viral Chemokine Inhibitors with Chemokines (pp. 167-180)
    Alcami, Antonio
14. Discovery of Small-Molecule Antagonists of Chemokine Receptors: Screening Strategy and Assays (pp. 181-188)
    Fuentes, Maria Elena
15. Visualization and Analysis of Adhesive Events in Brain Microvessels by Using Intravital Microscopy (pp. 189-198)
    Constantin, Gabriela
16. Animal Models to Study Chemokine Receptor Function: In Vivo Mouse Models of Allergic Airway Inflammation (pp. 199-210)
    Lloyd, Clare M.; Gutierrez-Ramos, Jose-Carlos
17. Assessing the Role of Multiple Phosphoinositide 3-Kinases in Chemokine Signaling: Use of Dominant Negative Mutants Controlled by a Tetracycline-Regulated Gene Expression System (pp. 211-222)
    Curnock, Adam P.; Sotsios, Yannis; Ward, Stephen G.
18. In Vitro and In Vivo Models to Study Chemokine Regulation of Angiogenesis (pp. 223-232)
    Bernardini, Giovanni; Ribatti, Domenico; Spinetti, Gaia; Morbidelli, Lucia; Ziche, Marina; Santoni, Angela; Capogrossi, Maurizio C.; Napolitano, Monica
19. Real-Time In Vitro Assay for Studying Chemoattractant- Triggered Leukocyte Transendothelial Migration Under Physiological Flow Conditions (pp. 233-242)
    Cinamon, Guy; Alon, Ronen
20. Generation of Monoclonal Antibodies Against Chemokine Receptors (pp. 243-260)
    Kremer, Leonor; Marquez, Gabriel
21. Detection of High-Affinity a4-Integrin Upon Leukocyte Stimulation by Chemoattractants or Chemokines (pp. 261-268)
    Chan, Jason R.; Cybulsky, Myron I.

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