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Cell migration is now well recognized as a critical component of the inflammatory disease process, so that its proper understanding promises to generate both ground-breaking basic discoveries and the development of novel therapeutics. In Cell Migration in Inflammation and Immunity: Methods and Protocols, leading cell biologists and immunologists present their most widely useful and innovative techniques for studying the molecular and cellular basis of this phenomenon. Describing each method in step-by-step detail, the authors provide a series of focused, cutting-edge techniques proceeding from the in vitro analysis of cell migration and the molecular mechanisms underlying this process, to methodologies for the analysis of cell migration in vivo. Methods for the analysis of rapid leukocyte adhesion under flow conditions in vitro are described, which may prove especially fruitful for scientists exploring the molecular mechanisms underlying both vascular recognition and leukocyte-endothelium interaction. Experimental approaches useful in establishing the role of cell migration in the pathogenesis of both acute and chronic inflammatory diseases are emphasized. Each fully tested protocol includes an introduction explaining the principle behind the technique, equipment and reagent lists, and tips on troubleshooting and how to avoid known pitfalls. Comprehensive and cutting-edge, Cell Migration in Inflammation and Immunity: Methods and Protocols offers novice and experienced investigators alike a collection of powerful techniques for studying the molecular basis and pathophysiological significance of cell migration in inflammatory and immune diseases, as well as for the development of novel therapeutics.
Immunology. --- Immunobiology --- Life sciences --- Serology
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Many of the genes found in the human major histocompatibility complex (MHC:HLA region) are subject to extreme polymorphism. HLA polymorphism has been used extensively in solid organ and bone marrow transplantation to match donors and recipients. Many MHC genes also have significant disease association and play a pivotal role in the control of immune response. In MHC Protocols, leading experts detail readily reproducible methods for identifying these polymorphisms in the human MHC. Described in step-by-step detail to ensure robust and successful experimental results, these techniques provide DNA-based protocols for the study of polymorphism in HLA class I and II genes and in non-HLA HMC genes associated with human disease (TAP1, TAP2, C2, C4, and TNF-a). The authors also describe methods for accessing HLA sequence data from electronic databases designed to catalog HLA-region genes and their alleles. Each method is written by an investigator who has used the technique extensively and includes troubleshooting tips and notes on pitfalls to avoid. Comprehensive and highly practical, MHC Protocols offers both experienced and novice investigators not only a state-of-the-art overview of this dramatically evolving area of immunology, but also a reliable guide to the identification of MHC gene polymorphisms.
Immunology. --- Immunobiology --- Life sciences --- Serology
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This extensively revised, updated, and expanded second edition of a classic laboratory manual applies new understanding of molecular mechanisms to the rational design and development of vaccines. Here the working researcher will find readily reproducible techniques for the development and production of live attenuated vaccines through the genetic manipulation of viruses and bacteria, vaccine vectors or inactivated toxins, and the production of synthetic peptides and conjugate vaccines. In addition, there are chapters describing more general techniques for vaccine formulation, delivery, and assessment of immune responses, and review discussions of scale-up to manufacture, vaccine quality assurance, and clinical trials. New to this second edition are cutting-edge chapters on two different viral vectors (Vaccinia and Semliki Forest), lactic acid bacteria for delivery of foreign antigens, incorporation of immunomodulators into plasmid DNA vaccines, construction of tetramer complexes to detect CMI responses, assessment of functional antibody responses, and the use of complete genome sequences for vaccine design. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes a background introduction outlining the principle behind the technique, equipment and reagent lists, tips on trouble shooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results. State-of-the-art and highly practical, Vaccine Protocols, Second Edition, provides essential information for those seeking to produce improved or novel vaccines in the rapidly evolving field of vaccine research and development.
Immunology. --- Immunobiology --- Life sciences --- Serology
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Superantigens, a class of proteins produced by bacteria and microbes, cause diseases ranging from food poisoning to toxic shock, but can also be used to understand these pathologies and devise therapeutic agents to combat them. In Superantigen Protocols, leading researchers in the biological, chemical, and physical investigation of superantigens describe in step-by-step detail their best experimental techniques to assess the physical characteristics and biological effects of these potent activators of the immune system. The protocols range from those for investigating the interactions of superantigens with cellular receptors to those for the analysis of their immunological and biological effects, including methods for using BIOcore to determine binding kinetics and establishing various lymphocyte cell culture systems. There are also accounts of such methods as the RNase protection assay, cytokine ELISA, FACS analysis, and cytokine production at the single cell level. Additional techniques for the discovery of new activators of the immune system, the development of new therapeutics for the suppression of superantigen-induced effects, and the treatment of inflammatory diseases are detailed. Comprehensive and cutting-edge, Superantigen Protocols provides a broad range of novel immunological and biochemical techniques that can be successfully applied not only to the structural, functional, and clinical properties of superantigens, but also to the many varieties of such proteins and to new cellular systems.
Immunology. --- Immunobiology --- Life sciences --- Serology
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Life sciences. --- Biosciences --- Sciences, Life --- Science
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Life Sciences --- Bioinformatics --- Genetics --- Genomics --- Human medicine
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