Full Description
During the last 30 years there has been a growing interest in cytokines as biological molecules able to regulate the most diverse functions in living org- isms, mainly at the level of cell-cell communication. Originally their definition was limited to the cells of the immune system (interleukins and lymphokines), but later that definition was extended to all cells, and their regulatory activity in such other processes as differentiation, apoptosis, angiogenesis, and wound he- ing has been now demonstrated. They comprise a group of small proteins (5-20 kDa) produced and released by cells in a tightly controlled fashion, active in the nano- or picomolar concentration range, and eliciting specific effects in nei- boring cells; therefore, their action is said to be autocrine, paracrine, or jux- crine. The latter property distinguishes them from hormones, which are produced by one tissue and are transported by the blood stream in order to act on a distant tissue. Chemokines are a subset of cytokines, but whether growth factors are included in the group is often a matter of discussion.
The activity of several cytokines can be inhibited by other cytokines or by biological response modi- ers; therefore, the latter are sometimes called "anti-cytokines. " The biological response of a particular cell is usually the result of the sum of all interactions with cytokines present at a certain time and in a certain sequence in time-the "cytokine network.
Contents
Large-Scale Generation of Plasmids that Express Type I Interferon.- Identification of trans-Acting Factors by Electrophoretic Mobility Shift Assay.- Modulation of the Interferon-? Signal by Transfection of Cells with an Antisense-RNA Expressing Vector.- Competitive RT-PCR to Quantify Small Amounts of mRNA.- In SituHybridization for Cytokines in Human Tissue Biopsies.- Purification, Identification, and Synthesis of Chemokines.- Receptor Isolation and Characterization.- Crystallization of Cytokine-Receptor Complexes.- Biosensor Analysis of Receptor-Ligand Interactions.- Analysis of SH2 Ligands and Identification of Sites of Interaction.- Assays for Antiviral Activity.- Assays for Cytotoxicity.- Assays for Chemotaxis.- In Vitro and In Vivo Assays for High-Molecular-Weight Pegylated Muteins of Granulocyte Colony-Stimulating Factor.- Development of a Mammalian Tet-On Expression Cell Line.- Immunohistochemical Detection of Cytokines in Human Tissue Sections.- Detection of Cytokine- and Chemokine-Expressing Cells at the Single Cell Level.- Detection of Cytokine Signal Transduction "Cross-Talk" in Leukocyte Activation.- Assay System for the Effect of Prostaglandin E2 in the Determination of Polarized Cytokine Production.
