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.

Luke O'Neill and Andrew Bowie bring together 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. 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.

This book presents key methodologies, tools and databases for biochemistry, microbiology and molecular biology in simple and straightforward language. Covering all aspects related to experimental principles and procedures, the protocols included here are brief and clearly defined, and include essential precautions to be taken while conducting experiments. The book is divided into two major sections: one on constructing, working with, and standard operating procedures for laboratory instruments; and one on practical procedures used in molecular biology, microbiology and biochemical analysis experiments, which are described in full. Each chapter describes both the basic theory and relevant practical details for a given experiment, and helps readers recognize both the experiment’s potential and limitations. Intended as an intensive introduction to the various tools used in molecular biology, the book covers all basic methods and equipment, including cloning, PCR, spectrophotometers, ELISA readers, sonicators, etc. As such, it offers a valuable asset for final year undergraduate (especially project) students, graduate research students, research scientists and technicians who wish to understand and employ new techniques in the field of biotechnology.

Immunotoxins represent a new class of human therapeutics that have widespread applications and a potential that has not yet been fully recognized since they were first conceived of by Paul Ehrlich in 1906. The majority of advances in the development and implementation of immunotoxins has occurred over the last 20 years. The reasons for this use of immunotoxins in basic science and clinical research are the powerful concurrent advances in genetic engineering and receptor physiology. Recombinant technology has allowed investigators to produce sufficient quantities of a homogeneous c- pound that allows clinical trials to be performed. The identification of specific receptors on malignant cell types has enabled scientists to generate immunotoxins that have had positive results in clinical trials. As more cellular targets are identified in coming years, additional trials will be conducted in different disease states affecting still larger patient populations. Modulation of the immune system to decrease the humoral response to immunotoxins may improve their overall efficacy. As increasingly more effective compounds are generated, it will be necessary to decrease the local and systemic toxicity - sociated with these agents, and methods for doing so are presently being - veloped. The work presented in Immunotoxin Methods and Protocols focuses on three specific areas of immunotoxin investigation that are being conducted by experts throughout the world. The first section describes the construction and development of a variety of immunotoxins.

Vaccine research and development is advancing at an unprecedented pace, with an increasing emphasis on rational design based upon a fundamental und- standing of the underlying molecular mechanisms. The aim of this volume is to provide a selection of contemporary protocols that will be useful to both novice and advanced practitioner alike. The variety of procedures required to design, develop, produce, and assess a vaccine is immense and covers aspects of ch- istry, biochemistry, molecular biology, cell biology, and immunology. No single volume can hope to cover these topics exclusively. Rather, here we attempt to provide a methods sourcebook focusing on hands-on practical advice. Comp- mentary and background information may be found in other volumes in the Me- ods in Molecular Medicine series. Of particular interest are volumes on Dendritic Cell Protocols, Interleukin Protocols, Vaccine Adjuvants, and DNA Vaccines. Since the publication of the first edition of Vaccine Protocols there have been major advances, particularly in the areas of bacterial genomics, antig- specific T-cell quantification, genetic manipulation of vaccine vectors, the h- nessing of natural molecules concerned with the regulation of immune responses, and the burgeoning field of DNA vaccinology. Hence, the extensive revision of this edition with new chapters on live viral vaccine vectors, atte- ated bacterial vectors, immunomodulators, MHC-peptide tetrameric complexes, and the identification of vaccine candidates by genomic analysis. Additionally, chapters from the first edition have been updated to accommodate state-of-t- art methods in vaccinology.

Interleukins in Cancer Biology responds to the growing need for credible and up-to-date information about the impact of interleukins on occurrence, development and progression of cancer. It provides reliable information about all known interleukins (38), describes recent discoveries in the field, and moreover, suggests further directions of research on the most promising aspects of this topic. The structure and presentation of the work is very understandable and clear with attention to detail maintained throughout. There are multiple illustrations throughout to help in comprehending and remembering the most important facts. . - Summarizes and discusses existing facts on the impact of all known interleukins in occurrence, development, and progression of cancer - Categorizes and clarifies all interleukins based on their role in cancer - Contains comprehensive and exhaustive information on each molecule

Cytokines are pleiotropic regulatory proteins involved in essentially all biological processes and associated with a wide variety of diseases, including inflammatory disorders as well as many types of cancer and leukemia. Knowledge about the quantitative and qualitative nature of cytokine production is critical in the understanding of normal and pathological processes. The cytokine detection in biological and clinical samples faces many challenges including their low abundance, the need to distinguish between active and latent cytokine forms, and the need to measure multiple cytokines in a single assay. This volume will provide a comprehensive collection of classic and cutting-edge methodologies that are currently used to analyze and quantify cytokines and their biological activities in complex biological and clinical samples. The chapters are divided into four main categories. The first group focuses on the immunodetection of released cytokines in tissue culture supernatants, plasma, serum and whole blood samples by immunoassays. These immunoassays measure the total concentrations of released cytokines regardless of their biological activity and include ELISA, flow cytometry, ELISPOT and the antibody-based proximity ligation. The second group will focus on the analysis of biologically active cytokines by bioassays using neutralizing antibodies, chemotaxis assay, cytokine-induced cell degranulation assay, cell proliferation and differentiation, cytokine-induced cytokine production and the radioreceptor cytokine assay. The third group focuses on the analysis of intracellular cytokines by flow cytometry, western blotting and fluorescence and confocal microscopy. In addition, this category includes protocols for quantitative analysis of cytokine gene expression by real time RT-PCR and analysis of the cytokine promoter occupancy by chromatin immunoprecipitation. The fourth group focuses on the recently developed multiplex arrays that can measure multiple cytokines in the same sample at the same time. This group includes quantification of multiple cytokines using cytometric bead arrays, ELISPOT assays, proteomics cytokine evaluation, multiplexed proximity ligation assays for high-throughput cytokine analysis and finally, cytokine gene expression analysis by gene arrays. The protocols will be written by experienced basic and clinical researchers with hands-on knowledge of the described protocols. By covering a broad variety of methods used in cytokine detection and analysis, this book will be of interest not only to biochemists, molecular biologists and immunologists but also to physician-scientists working in the field of cytokine research.