MHC Protocols

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.

Includes supplementary material: sn.pub/extras

Texte du rabat
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.

Contenu
Databases.- HLA Informatics.- Accessing HLA Sequencing Data Through the 6ace Database.- Polymorphism in Classical and Nonclassical Hla Genes.- HLA Typing by Restriction Fragment Length Polymorphism Analysis.- PCR-Restriction Fragment Length Polymorphism Typing of Class I and II Alleles.- PCR-Sequence-Specific Oligonucleotide Probe Typing for HLA-A, -B, and -DR.- HLA-DPA1 and -DPB1 Typing Using the PCR and Nonradioactive Sequence-Specific Oligonucleotide Probes.- PCR-Sequence-Specific Primer Typing of HLA Class I and Class II Alleles.- HLA Typing With Reference Strand-Mediated Conformation Analysis.- Sequencing Protocols for Detection of HLA Class I Polymorphism.- HLA-E and HLA-G Typing.- Typing Alleles of HLA-DM.- Polymorphism in Non-Hla Mhc Genes.- Typing Alleles of TAP1 and TAP2.- Determining Alleles of the C2 Gene by Southern Blotting.- Complement C4 Protein and DNA Typing Methods.- Typing of Tumor Necrosis Factor Alleles.- Molecular Typing of the MHC Class I Chain-Related Gene Locus.- Microsatellites.- HLA Microsatellite Analysis.