Cell Migration in Inflammation and Immunity Methods and Protocols

Chemokines and their receptors play a central role in the pathogenesis of numerous, perhaps all, acute and chronic inflammatory diseases. About 50 distinct chemokines produced by a variety cell types and tissues either c- stitutively or in response to inflammatory stimuli are involved in a plethora of biological processes. These small secreted proteins exert their exquisitely variegated functions upon binding to a family of seven-transmembrane spanning G-protein coupled receptors (GPCRs) composed of almost 20 distinct entities. The biological activities of chemokines range from the control of leukocyte trafficking in basal and inflammatory conditions to the regulation of hema- poiesis, angiogenesis, tissue architecture, and organogenesis. The basis for such diversified activities rests, on one hand, upon the ubiquitous nature of chemokine production and chemokine receptor expression. Virtually every cell type can produce chemokines and expresses a unique combination of chemokine receptors. On the other hand, chemokine receptors make use of a flexible and complex network of intracellular signaling machineries that can regulate a variety of cellular functions ranging from cell migration, growth, and differentiation to death. As knowledge of the size of chemokine and chemokine receptor families rapidly reaches completeness, much is still to be uncovered in terms of fu- tional architecture of the chemokine system. The disparity between the large number of chemokines and that smaller number of receptors is balanced by the promiscuity in ligand–receptor interactions, with multiple chemokines binding to the same receptor and several chemokines binding to more than one receptor.

Chemotaxis and Interaction with Vascular or Lymphatic Endothelium Silvano Sozzani, Annunciata Vecchi, Paola Allavena, and Alberto Mantovani Analysis of Integrin-Dependent Rapid Adhesion Under Laminar-Flow Conditions Carlo Laudanna Posttranslational Processing of Chemokines Paul Proost, Frank Mahieu, Evemie Schutyser, and Jo Van Damme Chemotactic Profiling of Lymphocyte Subpopulations Lucia Colantonio, Andrea Iellem, and Daniele D'Ambrosio Measurement of the Levels of Polymerized Actin (F-Actin) in Chemokine-Stimulated Lymphocytes and GFP-Coupled cDNA Transfected Lymphoid Cells by Flow Cytometry Miguel Vicente-Manzanares, Mariano Vitón, and Francisco Sánchez-Madrid Evaluation of Rho Family Small G-Protein Activity Induced by Integrin Ligation on Human Leukocytes Angela Gismondi, Fabrizio Mainiero, and Angela Santoni Reconstructing Leukocyte Migration in 3D Extracellular Matrix by Time-Lapse Videomicroscopy and Computer-Assisted Tracking Peter Friedl and Eva-B. Bröcker Analyzing Chemotaxis Using Dictyostelium discoideum as a Model System Mark A. Landree and Peter N. Devreotes Conditional Transgenic Models to Study Chemokine Biology Sergio A. Lira, Borna Mehrad, Shu-Cheng Chen, Petronio Zalamea, David J. Kinsley, Maria T. Wiekowski, Elizabeth Coronel, Galya Vassileva, Denise Manfra, and Kristian K. Jensen Intravital Microscopy as a Tool for Studying Recruitment and Chemotaxis Denise C. Cara and Paul Kubes Tracking Antigen-Specific Lymphocytes In Vivo Claire L. Adams, Catherine M. Rush, Karen M. Smith, and Paul Garside Analysis of Homing-Receptor Expression on Infiltrating Leukocytes in Disease States Margherita Mariani and Paola Panina-Bordignon Interaction of Viral Chemokine Inhibitors with Chemokines Antonio Alcami Discovery of Small-Molecule Antagonists ofChemokine Receptors: Screening Strategy and Assays Maria Elena Fuentes Visualization and Analysis of Adhesive Events in Brain Microvessels by Using Intravital Microscopy Gabriela Constantin Animal Models to Study Chemokine Receptor Function In Vivo: Mouse Models of Allergic Airway Inflammation Clare M. Lloyd and Jose-Carlos Gutierrez-Ramos Assessing the Role of Multiple Phosphoinositide 3-Kinases in Chemokine Signaling: Use of Dominant Negative Mutants Controlled by a Tetracycline-Regulated Gene Expression System Adam P. Curnock, Yannis Sotsios, and Stephen G. Ward In Vitro and In Vivo Models to Study Chemokine Regulation of Angiogenesis Giovanni Bernardini, Domenico Ribatti, Gaia Spinetti, Lucia Morbidelli, Marina Ziche, Angela Santoni, Maurizio C. Capogrossi, and Monica Napolitano Real Time In Vitro Assay for Studying Chemoattractant-Triggered Leukocyte Transendothelial Migration Under Physiological Flow Conditions Guy Cinamon and Ronen Alon Generation of Monoclonal Antibodies Against Chemokine Receptors Leonor Kremer and Gabriel Márquez Detection of High-Affinity a4-Integrin Upon Leukocyte Stimulation by Chemoattractants or Chemokines Jason R. Chan and Myron I. Cybulsky Index