Chemokine Protocols

The chemokines family of small proteins are involved in numerous b- logical processes ranging from hematopoiesis, angiogenesis, and basal l- kocyte trafficking to the extravasation and tissue infiltration of leukocytes in response to inflammatory agents, tissue damage, and bacterial or viral infection. Chemokines exert their effects through a family of seven G-protein coupled transmembrane receptors. Worldwide interest in the chemokine field surged dramatically early in 1996, with the finding that certain chemokine receptors were the elusive coreceptors, required along with CD4, for HIV infection. Today, though over 40 human chemokines have been described, the n- ber of chemokine receptors lags behindonly 17 human chemokine receptors have been identified so far. What has emerged over the years is that most chemokine receptors bind several distinct ligands, and indeed the majority of chemokines are able to bind to multiple chemokine receptors, explaining to some extent the apparent disparity in the numbers of chemokines and rec- tors. Yet in spite of the apparent redundancy in chemokine/chemokine rec- tor interactions, it is clear that in vivo, spatial, temporal, and indeed cell- and tissue-specific expression of both chemokines and their receptors are imp- tant factors in determining the precise nature of cellular infiltrates in phy- ological and pathological processes.

Includes supplementary material: sn.pub/extras

Zusammenfassung
highly recommended...Seldom does on find so much valuable information, concisely presented and with such obvious potential for the future, made available in one volume." - E-Streams

"...Chemokine Protocols is a collection of clear, well written protocols and I would recommend it as an excellent starting point for any scientist with an interest in chemokines." - Molecular Biotechnology

Inhalt
Purification of Chemokines from Natural Sources.- Cloning of Novel Chemokines Using a Signal Sequence Trap Method.- Chemokine Expression in Insect Cells.- Expression of Chemokines in Escherichia coli.- Expression of Chemokines in the Periplasmic Space of E. coli.- Synthesis of Chemokines.- Identification of Novel Chemokines From Expressed Sequence Tag Databases.- Purification of Recombinant Chemokines from E. coli.- Chemokine Receptor Cloning.- Generation of Stable Cell Lines Expressing Chemokine Receptors.- Modified Microchemotaxis Assays.- Transwell Chemotaxis.- Endothelial Cell Chemotaxis Assays.- Radiolabeled Chemokine Binding Assays.- Scintillation Proximity Binding Assay.- Calcium Mobilization.- Actin Polymerization.- Reactive Oxygen Release.- Histamine Release.- Measurement of Phosphoinositide 3-Kinase Activity.- Glycosaminoglycan Binding Assays.- CFU-A Assay for Measurement of the Antiproliferative Effects of Chemokines on Murine Early Hemopoietic Progenitors.- Downmodulation and Recycling of Chemokine Receptors.- Analysis of Chemokine Receptor Endocytosis and Recycling.- Chemokine Inhibition of HIV Infection.- 26The Production of Chemokine Specific Monoclonal Antibodies.- Monoclonal Antibodies to Chemokine Receptors.- Targeted Expression of Chemokines In vivo.- Chemokine Knockout Mice.- Chemokine Receptor Knockout Mice.- Measurement of Eosinophil Accumulation In vivo.- Murine Model of Allergic Lung Inflammation.- Murine Models of Airway Inflammation.- Rat Models of Respiratory Inflammation.- Murine Model of Crescentic Nephritis.- Rabbit Models of Pneumonia, Peritoneal Sepsis, and Lung Injury.