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.

Antigen processing and presentation, as a field, explores a broad range of protein interactions and functions, both intracellular (in the cytoplasm and in the endoplasmic reticulum) and at the cell surface (between T cells and MHC molecules). To investigate such a diverse array, it is necessary that biochemical, cell biology, and immunological techniques all be employed. The purpose of Antigen Processing and Presentation Protocols is therefore to detail the most up-to-date techniques being used in this burgeoning field. Such techniques include those used 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 MHC/peptide. Antigen Processing and Presentation Protocols should aid both those new and those experienced in this area of research in extending the questions that can be asked and answered by the application of these current methods. For editorial assistance, I would like to thank Angela Beninga and Rachael Turnquist.

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.

Recently expanded to 2 volumes, Short Protocols in Molecular Biology, Fifth Edition, provides condensed descriptions of more than 700 methods compiled from Current Protocols in Molecular Biology. Includes new chapters on chromatin assembly and analysis, nucleic acid arrays, generation and use of combinatorial libraries, discovery and analysis of differentially expressed genes in single cells and cell populations. The book is specifically designed to provide quick access to step-by-step instructions for the essential methods used in every major area of molecular biological research Short Protocols in Molecular Biology, Fifth Edition is an authoritative and indispensable guide for all life scientists, researchers, and students at the graduate and advanced undergraduate level Expanded to 2 volumes.