Induction of epitope-specific cytotoxic T lymphocyte (CTL) responses can improve the outcome of disease in a variety of model systems. However, natural immunity or that arising by immunization with whole proteins or organisms with multiple potential epitopes, may not always result in t ...
Microarray experiments rely on and generate (1–3) quantities of data that are simply too large to be stored on a researcher’s desktop computer in spreadsheet format. A single microarray may have 40,000 spots. Associated with each spot may be as many as 40 different metrics, and an experimental seri ...
DNA microarray technology has been used successfully to detect the expression of many thousands of genes, to detect DNA polymorphisms, and to map genomic DNA clones (1–4). It permits quantitative analysis of RNAs transcribed from both known and unknown genes and allows one to compare gene expr ...
Methods to transiently and stably transfect blood stages of the human malaria parasite Plasmodium falciparum have been developed and adapted for gene-knockout, allelic replacement, and transgene expression in this organism. These methods are detailed in this chapter, as are appro ...
The presence of preformed donor-specific HLA antibodies detected by Complement-dependent cyto�toxicity (CDC) crossmatch assay is associated with a high incidence of hyperacute or accelerated rejection and remains one of the gold standard tests pre-transplant. The standard CDC ...
Antibodies against MICA have been found in organ transplant recipients and were found to be associated with decreased survival of kidney allografts. The MICA antibody screening assay is a Luminex-based solid phase immunoassay designed to detect IgG antibodies binding to beads pre-co ...
Solid phase immunoassays for the detection and characterization of HLA-specific antibodies provide greatly increased sensitivity, specificity, and time and reagent efficiency, compared to the traditionally used cell-based methods. Testing is performed using commercia ...
The ability to directly measure virus-specific lymphocytes using fluorochrome-labeled tetrameric complexes has proven a great advancement for the transplantation field. Viral peptide/HLA tetrameric complexes allow the rapid generation of virus-specific clones using ...
T cell alloreactivity is responsible for much of the morbidity and mortality associated with tissue transplantation and graft versus host disease. Immunoassays for ex vivo monitoring and quantitation of alloreactive T cells are being increasingly utilised to provide valuable in ...
The major histocompatibility complex (MHC) on chromosome 6 is one of the most intensively studied regions of the human genome and has many features which make it unique. It is the source of much research interest because of its role in autoimmune and infectious disease susceptibility, and of diag ...
Spleen and lymph node retrieved post-mortem from deceased organ donors are a rich source of lymphocytes. Storage of lymphocytes separated from these sources can be valuable where post-transplant testing (crossmatching) is required. DNA extraction from stored lymphocytes also al ...
Natural killer (NK) cells are more than simple killers and have been implicated in control and clearance of malignant and virally infected cells, regulation of adaptive immune responses, rejection of bone marrow transplants, and autoimmunity and the maintenance of pregnancy. Human NK c ...
The flow cytometric lymphocyte crossmatch is a standard technique for evaluating the compatibility of potential kidney transplant recipients and donors. Recipient serum is incubated with donor lymphocytes and the latter are analysed in a flow cytometer for the presence of bound IgG a ...
The killer-cell immunoglobulin-like receptors (KIR), which enable NK cells to detect allogeneic target cells and abnormalities in the expression of self-HLA molecules, are encoded by genes that display extensive copy number variation. These variations in the KIR genotype are relev ...
The Collaborative Transplant Study (CTS) was initiated in 1982 in Heidelberg, Germany, and originated from the need to gain further insight into the complex problems and risks involved in human organ transplantation. Currently, more than 400 transplant centers in 45 countries are contr ...
Disease association studies involving highly polymorphic immunogenetic data may involve analyses at one or many units of analysis, including amino acid, allele, genotype and haplotype levels, as well as consideration of gene–gene or gene–environment interactions. The selecti ...
In this chapter, we describe analyses commonly applied to immunogenetic population data, along with software tools that are currently available to perform those analyses. Where possible, we focus on tools that have been developed specifically for the analysis of highly polymorphic im ...
In this chapter, we outline some basic principles for the consistent management of immunogenetic data. These include the preparation of a single master data file that can serve as the basis for all subsequent analyses, a focus on the quality and homogeneity of the data to be analyzed, the documentat ...
We have developed a MICA typing method based on polymerase chain reaction (PCR) sequence-based typing and a computer program that determines the polymorphisms and distinguishes the GCT repeats in exon 5. One PCR amplification was performed to obtain templates of 2.2 kb, including exons 2, 3, 4, and 5 ...
Real-time quantitative PCR is an efficient method for high-throughput genotyping of single nucleotide polymorphisms (SNPs). In this chapter, we describe the 5′ nuclease allelic discrimination assay for genotyping biallelic SNPs.