Nucleotide-binding domain leucine-rich-repeat containing receptors; NOD-Like Receptors (NLRs) were originally described as microbial sensors involved in host defense against pathogens that comprise an important component of the innate immune system. Recently, their ce ...

Through its ability to control the proteolytic maturation and secretion of interleukin-1 family cytokines, the inflammasome occupies a central role in the activation of inflammation and also influences the shaping of adaptive immunity. Since it affects a multitude of different imm ...

DNA sequencing is a powerful technique for identifying allelic variation within the human leukocyte antigen (HLA) genes. Sequencing is usually focused on the most polymorphic exons of the class I (HLA-A, -B, -C) and class II (HLA-DR, -DQ, and -DP) genes. These exons encode the antigen recognition s ...

HLA typing by sequence-specific primers (PCR-SSP) is a commonly used technique in HLA typing in which multiple pairs of cis-located allele-specific primers are used to determine the alleles present in a given DNA sample. Although the technique is around two decades old, it still offers a relati ...

Transcriptome analysis or global gene expression profiling is a powerful tool for discovery as well as �understanding biological mechanisms in health and disease. We present in this chapter a description of methods used to isolate mRNA from cells and tissues that has been optimized for pre ...

Because of our access to human genome data and ever improving genome sequencing and proteome analysis methods we are much better in terms of our understanding of biological processes. In addition to genomics, proteomics, and other “omics” methods, availability of more sophisticated mol ...

Tolerogenic antigen presenting cells (APC), primarily dendritic cells (DC), are essential to the induction and maintenance of immunologic tolerance in clinical transplantation. They induce the differentiation of CD8+ T suppressor (Ts) and CD4+ T regulatory (Treg) or anergic cells, w ...

The intracellular cytokine method allows for a multiparametric readout by flow cytometry with precise phenotyping of the responding T cells. The intracellular cytokine staining of cells that have been fixed and permeabilized following a short-term activation and staining with an ...

The use of immunosuppression in solid organ transplantation is associated with increased morbidity and mortality. Monitoring trough immunosuppression levels alone in transplant recipients is insufficient to gauge the suppression of immune responses. The ImmuKnow Immune C ...

The enzyme-linked immunosorbent assay (ELISA) is a widely used technique for detecting antibodies (Abs) and is employed in clinical laboratories to identify Abs against various self-antigens–autoAb development and quantitation. This method relies on specific antigen–Ab int ...

The detection of auto-antibodies detection is a major aspect of patient’s immunomonitoring. Antibodies directed against the heart and kidney Angiotensin II type 1 receptor (AT1R) play a major role in antibody mediated rejection after heart and kidney transplantation and in oblitera ...

Flow cytometric crossmatch tests provide a donor-specific, cell based method for the detection of alloreactive antibodies in the sera of transplant patients. Conventional crossmatch tests used in solid organ transplantation utilize lymphocytes as target cells to detect the pre ...

HLA-specific B cells can be identified, quantified, and isolated after staining with HLA tetramers. Quantification of these B cells can in turn identify individuals who are sensitized to HLA antigens and the isolation of these cells facilitates a variety of experimental investigation ...

Solid phase Luminex� and flow cytometric single antigen bead assays offer exquisite sensitivity and specificity for HLA antibody detection. Unlike the historical complement-dependent cytotoxicity (CDC) method, these assays do not distinguish complement fixing from non-co ...

The definition of HLA-specific antibodies in solid organ transplant patients is a necessary tool for recipient selection prior to transplantation and monitoring for rejection post transplant. Solid phase assays can detect both complement fixing and non-complement fixing HLA-s ...

Flow cytometry is an invaluable tool for studying lymphocyte biology. In transplantation, flow cytometry represents the most sensitive method for demonstrating the binding of HLA antibodies to cellular targets. Referred to as the flow cytometric crossmatch (FCXM), the implement ...

The complement-dependent cytotoxic crossmatch is an informative test that detects alloantibodies in pre- and post-transplant patients, which may dictate clinical management of transplant patients. While challenging to perform, the cytotoxic crossmatch represents the on ...

By interacting with specific HLA class I molecules, the killer cell immunoglobulin-like receptors (KIR) regulate the effector function of natural killer (NK) cells and subsets of CD8 T cells. The KIR receptors and HLA class I ligands are encoded by unlinked polymorphic gene families located ...

Engraftment monitoring is critical for patients after Hematopoietic Stem Cell Transplantation (HSCT). Complete donor chimerism is the goal; therefore, early detection of rejection and relapse is crucial for guiding the patient post HSCT treatment. Quantitative PCR for chimeri ...

Next-generation sequencing (NGS) of HLA class I and II loci (HLA-A, HLA-B, HLA-C, DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, DPB1) is described here in detail using the 454 Life Sciences GS FLX System and Titanium chemistry. An overview of the protocol with our experience on sequence performance efficiencies, ...