Interleukins are a family of proteins that regulate the maturation, diff- entiation, or activation of cells involved in immunity and inflammation, and belong to a broader family termed cytokines. Collectively these proteins are the key orchestrators of host defense and the response to tissue injury. There are currently 23 different interleukins (numbered from IL-1 to IL-23), although the full extent of the interleukin family will only become clear upon analysis of the human genome sequence. Most important, interleukins are central to the pathogenesis of a wide range of diseases that involve an immune com- nent, including such conditions as rheumatoid arthritis, multiple sclerosis, ulcerative colitis, psoriasis, and asthma. Interleukins have also been imp- cated in other conditions, including cancer, migraine, myocardial infarction, and depression. In essence, when cells are activated by interleukins, a program of gene expression is initiated in the target cell that alters the cell's phenotype, leading to enhanced immune reactivity, inflammation, and/or proliferation. Interleukins are therefore at the core of the cellular basis for many diseases. They are the subject of intense investigation by biomedical researchers and the targeting or use of interleukins in the clinic is proceeding apace. Approaches such as t- geting IL-4 in asthma or IL-1 in joint disease are being pursued, and it is likely that in the next 510 years a number of new therapies based on either inhib- ing or administering interleukins will be available.

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Klappentext
Interleukins, a family of proteins that regulate immunity and inflammation throughout the body, are also involved in autoimmune disease (rheumatoid arthritis, multiple sclerosis, and asthma); are implicated in such other conditions as cancer, myocardial infarction, and depression; and have value in the diagnosis and prognosis of many diseases. Luke O'Neill and Andrew Bowie bring together in Interleukin Protocols a collection of standard and advanced methods for measuring these powerful agents. Their readily reproducible techniques range from the assay of interleukin protein and mRNA-using ELISA, FACS, and RT-PCR-to the study of interleukin signal transduction. Newer techniques are also covered, including the analysis of interleukin gene polymorphisms and the use of cDNA microarrays. Many of the assays are geared to specific pathologies, including breast cancer, depression, psoriasis, Grave's disease, migraine, and myocardial infarction. There are also helpful discussions of the difficulties in measuring interleukins in different biological fluids, such as sputum from asthma patients, peritoneal fluids, synovial fluid from arthritic joints, and cerbrospinal fluid from patients with meningitis. Each step-by-step method is described by a hands-on expert and includes notes on how to avoid failure.
Comprehensive and highly practical, Interleukin Protocols offers biomedical investigators a stellar collection of all the major techniques needed to analyze the role of interleukins in disease, to improve diagnosis, and to foster the rapid emergence of new and more powerful therapeutics.

Inhalt
General Methods for Assaying Interleukin Protein and mRNA.- ELISAs and Interleukin Research.- Elispot Technique for Assaying Interleukins.- Measurement of Interleukins by ELISPOT Assay with Particular Application to Dual-Color Analysis (Stardust Assay).- Assaying Interleukins Intracellularly by FACS.- Basic RT-PCR for Measurement of Cytokine Expression.- Quantitative PCR for Measurement of Cytokine Expression.- Single Cell RT-PCR for the Detection of Cytokine mRNA.- Quantitation of Cytokine mRNA by Flash-Type Bioluminescence.- Measurement of Interleukin mRNA by Northern Blotting Using a Nonradioactive Detection Method.- Quantification of Mouse IL-6 and TNF- ? mRNA Using Xplore Assays.- Measurement of Cytokine and Chemokine mRNA Using Nonisotopic Multiprobe RNase Protection Assay.- Isolation of Total and Bioactive Interleukins by Immunoaffinity-Receptor Affinity Chromatography.- Detection of Cytokines by Immunohistochemistry.- Whole Blood Assays and the Influence of Circadian Rhythmicity on Human Cytokine Measurement.- Illustration of Methodologies Using Individual Interleukins.- Assaying Interleukin-8.- Measurement of Interleukin-2 by Bioassay.- Assaying Interleukin-15.- Assaying Interleukins In Particular Pathologies.- Assaying Interleukin-6 in Breast Cancer.- Cytokines, Stress, and Depressive Illness.- Measurement of Interleukins in Cutaneous Disorders.- Interleukins in Graves? Disease.- Evaluation of Cytokines in Migraine Patients by ELISA.- Assaying Interleukins in Plasma in the Course of Acute Myocardial Infarction.- Assaying Interleukins in Particular Biological Fluids.- Measurement of Cytokines in Peritoneal Fluids.- Measurement of Cytokines in Seminal Plasma.- Measurement of Cytokines in Induced Sputum.- Cytokines in Blister Fluids.- Measurement of Cytokinesin Synovial Fluid.- Measurement of Cytokines in Cerebrospinal Fluid.- Detection of Cytokines in Tears.- Newer Techniques: Interleukin Signal Transduction,.- Measurement of Phosphoinositide 3-Kinase Activation by Interleukins.- Analysis of Interleukin-2 Signaling Using Affinity Precipitations and Polyacrylamide Gel Electrophoresis.- Applications for Green Fluorescent Protein in Cell Signaling.- Use of Pichia pastoris for Production of Recombinant Cytokines.- 35Analysis of Single Nucleotide Polymorphisms in the Interleukin-1B Gene Using 5? Nuclease Assays.- Measurement of Cytokine and Chemokine Gene Expression Patterns Using cDNA Array.