O’Neill / Bowie Interleukin Protocols

Erscheinungsjahr 2008
ISBN: 978-1-59259-146-6
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 60, 443 Seiten, eBook

Reihe: Methods in Molecular Medicine

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 5–10 years a number of new therapies based on either inhib- ing or administering interleukins will be available.

Zielgruppe

Research

Autoren/Hrsg.

O’Neill, Luke A. J.

Bowie, Andrew

Inhaltsverzeichnis

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.