Humana Cell Migration in Inflammation and Immunity: Methods and Protocols: 239 (Methods in Molecular Biology, 239)

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. From the Back Cover Cell migration is now well recognized as a critical component of the inflammatory disease process, so that its proper understanding promises to generate both ground-breaking basic discoveries and the development of novel therapeutics. In Cell Migration in Inflammation and Immunity: Methods and Protocols, leading cell biologists and immunologists present their most widely useful and innovative techniques for studying the molecular and cellular basis of this phenomenon. Describing each method in step-by-step detail, 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. 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 both vascular recognition and leukocyte-endothelium interaction. Experimental approaches useful in establishing the role of cell migration in the pathogenesis of both acute and chronic inflammatory diseases are emphasized. 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. Comprehensive and cutting-edge, Cell Migration in Inflammation and Immunity: Methods and Protocols offers novice and experienced investigators alike a collection of powerful techniques for studying the molecular basis and pathophysiological significance of cell migration in inflammatory and immune diseases, as well as for the development of novel therapeutics.