A collection by leading experts provide readily reproducible methods for identifying 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, TAP 2, C2, C4, and TNF-). The authors also describe methods for accessing HLA sequence data from electronic databases designed to catalog HLA-region genes and their alleles. Use state-of-the-art methods to identify MHC gene polymorphisms Access HLA sequence data from electronic databases.
The processing and presentation of antigens by major histocompatibility complex (MHC) molecules is a crucial immunological phenomenon in infectious disease, malignancy, autoimmune disease, and transplantation. In Antigen Processing and Presentation Protocols, well-recognized and innovative experimentalists detail their cutting-edge methods for studying this complex process. Drawing on expertise from biochemistry, cell biology, and immunology, they describe step-by-step methods designed to question how MHC-binding peptides are generated, to test how peptides are delivered to MHC molecules, to analyze MHC peptide-binding patterns, and to assay the T cell response to the MHC/peptide complex. Each method is presented in sufficient detail to be readily reproducible and includes notes about potential pitfalls and tips on how to avoid failures. Emphasis is given those technical steps critical for experimental success that are often omitted from methods published in the primary literature. Eminently accessible and state-of-the-art, Antigen Processing and Presentation Protocols provides both new and experienced investigators with highly practical tools that will extend the questions that can be asked, and effectively be answered, concerning antigen processing/presentation.
Transplantation Immunology Edited by Fritz H. Bach, M.D., and Hugh Auchincloss, Jr., M.D. In recent years, transplantation immunology has evolved as a distinct field founded on the recognition that rejection of a transplanted organ or tissue is mediated by immune mechanisms in the host responding to antigens in the donor tissue. This book provides a comprehensive, up-to-date review of current knowledge regarding the specific immune mechanisms which can cause recipients to reject clinical transplants. The text is divided into four sections: Major and Minor Histocompatibility Antigens, Immunobiology of Graft Rejection, Clinical Transplantation, and Frontiers in Transplantation. The first two sections cover present-day understanding of the genetic controls of immunity as well as the two basic concerns of transplantation immunology, namely donor tissue antigens and the host cellular response. The third section deals with clinical aspects of transplantation, while the fourth looks toward future efforts of scientific research in this field—including advances in tolerance and xenotransplantation. A detailed appendix entitled "Molecular Biology for the Clinician" is included to help make the material accessible to readers of all backgrounds. Authored by a wide range of experts in both clinical and research fields, individual chapters cover such topics as: T cell immunity Antigen presentation Alloreactivity Pancreas and islet cell transplantation Allogeneic bone marrow transplantation Models of tolerance induction Xenotransplantation The swine leukocyte antigen (SLA) complex Transplantation Immunology is an ideal teaching text for the fundamental concepts, latest findings, and future directions in this increasingly important field. This volume will appeal to a wide range of clinicians including surgeons and primary-care physicians, as well as scientists in molecular biology and immunology.
From the simple discovery in 1962 that resorbing tadpole tail expressed an enzyme (MMP) that could degrade collagen gels, matrix metalloproteinase (MMP) research has advanced to discover more than twenty distinct vertebrate MMPs and four specific inhibitors (TIMPS), a veritable family of enzymes involved in many physiological and pathological processes. In Matrix Metalloproteinase Protocols, leading experts detail proven laboratory techniques for the study of MMPs. The methods include those for the expression and purification of MMPs and TIMPs, for the detection of MMPs and TIMPs at both the protein and mRNA levels, and for the assay of MMP and TIMP activities in a wide variety of circumstances. Each method includes step-by-step instructions and notes on variant applications and pitfalls to avoid. A selective overview of the MMP arena spells out where the field has been, where it is, and where it is going. Comprehensive and highly practical, Matrix Metalloproteinase Protocols brings together the long and hard-earned experience of master experimentalists that will allow not only novices to get up to speed quickly, but also add to the repertoire of successful techniques in expert laboratories.
The aim of MHC Protocols is to document protocols that can be used for the analysis of genetic variation within the human major histocompatibility complex (MHC; HLA region). The human MHC encompasses approximately 4 million base pairs on the short arm of chromosome 6 at cytogenetic location 6p21. 3. The region is divided into three subregions. The telomeric class I region contains the genes that encode the HLA class I molecules HLA-A, -B, and -C. The centromeric class II region contains the genes encoding the HLA class II molecules HLA-DR, -DQ, and -DP. In between is the class III region, originally identified because it contains genes encoding components of the complement pathway. The entire human MHC has recently been sequenced (1) and each subregion is now known to contain many other genes, a number of which have immunological functions. The study of polymorphism within the MHC is well established, because the region contains the highly polymorphic HLA genes. HLA polymorphism has been used extensively in solid organ and bone marrow transplantation to match donors and recipients. As a result, large numbers of HLA alleles have been identified, a process that has been further driven by recent interest in HLA gene diversity in ethnic populations. The extreme genetic variation in HLA genes is believed to have been driven by the evolutionary response to infectious agents, but relatively few studies have analyzed associations between HLA genetic variation and infectious disease, which has been difficult to demonstrate.
The second edition volume expands on the previous edition with new and updated chapters on the latest methods used for studying presentation of antigenic peptides produced in the standard processing pathways for MHC class I and II molecules. The new chapters cover topics such as biochemical and cellular approaches to study the impact of the endoplasmic reticulum aminopeptidases; techniques to monitor MHC class I synthesis and degradation; approaches to measure processing efficacy; description of different assays measuring MHC recycling; and protocols to produce MHC class II tetramers. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, Antigen Processing: Methods and Protocols, Second Edition is a valuable tool for both novice and expert researchers interested in studying antigen processing and venturing out further into this evolving field.
Current Protocols in Immunology is a three-volume looseleaf manual that provides comprehensive coverage of immunological methods from classic to the most cutting edge, including antibody detection and preparation, assays for functional activities of mouse and human cells involved in immune responses, assays for cytokines and their receptors, isolation and analysis of proteins and peptides, biochemistry of cell activation, molecular immunology, and animal models of autoimmune and inflammatory diseases. Carefully edited, step-by-step protocols replete with material lists, expert commentaries, and safety and troubleshooting tips ensure that you can duplicate the experimental results in your own laboratory. Bimonthly updates, which are filed into the looseleaf, keep the set current with the latest developments in immunology methods. The initial purchase includes one year of updates and then subscribers may renew their annual subscriptions. Current Protocols publishes a family of laboratory manuals for bioscientists, including Molecular Biology, Human Genetics, Protein Science, Cytometry, Cell Biology, Neuroscience, Pharmacology, and Toxicology.
Abnormal expression of MHC class I molecules in malignant cells is a frequent occurrence that ranges from total loss of all class I antigens to partial loss of MHC specific haplotypes or alleles. Different mechanisms are described to be responsible for these alterations, requiring different therapeutic approaches. A complete characterization of these molecular defects is important for improvement of the strategies for the selection and follow-up of patients undergoing T-cell based cancer immunotherapy. Precise identification of the mechanism leading to MHC class I defects will help to develop new personalized patient-tailored treatment protocols. There is significant new research on the prevalence of various patterns of MHC class I defects and the underlying molecular mechanisms in different types of cancer. In contrast, few data is available on the changes in MHC class I expression during the course of cancer immunotherapy, but the authors have recently made discoveries that show the progression or regression of a tumor lesion in cancer patients undergoing immunotherapy depends on the molecular mechanism responsible for the MHC class I alteration and not on the type of immunotherapy used. According to this notion, the nature of the preexisting MHC class I lesion in the cancer cell has a crucial impact on determining the final outcome of cancer immunotherapy. This SpringerBrief will present how MHC class 1 is expressed, explain its role in tumor progression, and its role in resistance to immunotherapy.
In Gene Therapy Protocols, Volumes 1 and 2, internationally recognized investigators describe cutting-edge laboratory techniques for the study of Production and In Vivo Applications of Gene Transfer Vectors (Volume 1) and Design and Characterization of Gene Transfer Vectors (Volume 2). In this second volume, readers will find a comprehensive resource of current and emerging methods for the processing and characterization of viral and non-viral gene transfer vectors.
This publication provides an update on the current status of gene maps in different livestock and pet/companion animal species. The findings summarized in species specific commentaries and original articles testify the rapid advances made in the field of animal genomics. Of significant interest is the fact that current investigations are providing headways for two important and exciting research fronts: targeted high-resolution mapping leading to the application of genomic information in addressing questions of economic and biological significance in animals, and the initiation of whole genome sequencing projects for some of the animal species. Like in humans and mice, this will set the stage for a new level of research and real time complex analysis of the genomes of these species. Animal Genomics signifies the beginning of a new era in this field and celebrates the achievements of the past 20 years of genomics research. It will be of special interest to researchers involved in genome analysis - both gross chromosomal as well as molecular - in various animal species, and to comparative and evolutionary geneticists.