免疫学技术

Since NK cells specialize in contact-dependent functions including cytotoxicity, interest has focused on the direct study of the interface between the NK cell and the cell with which it is interacting. This interface is also known as the immunological synapse and is characterized by an extr ...

Although NK cells were originally identified as “naturally” active cells believed to follow a cell-autonomous activation program, it is now widely accepted that NK cells need to interact with dendritic cells for their full functional activation and for their homeostasis. In this chapte ...

This flow cytometry-based method is a quick way to detect adhesion of NK cells to target cells. The two cell types are labeled with distinct fluorescent dyes and following co-incubation, the number of NK cells firmly adhered to target cells is quantified using two-color flow cytometry.

Determining the contribution of individual receptors to natural killer (NK) cell function is complicated by the multiplicity of activating and inhibitory NK cell receptors. Mammalian target cells typically express a variety of ligands for NK cell receptors. Engagement of NK cell rec ...

Although the use of mutant mice has been extremely useful in identifying those proteins and molecules specifically required for the development of NK cells, the establishment of a well-defined protocol to replicate in vitro the major steps corresponding to the process of NK cell different ...

Natural killer (NK) cells express receptors to detect and kill target cells based on expression of target cell surface molecules. Through a process termed NK cell licensing, only NK cells that express inhibitory receptors (e.g., Ly49 receptors in the mouse) for self-major histocompatibil ...

Identification of natural killer (NK) cell subsets has gained attention with the recent discovery that mature mouse NK cells comprise two distinct stages. Delineation of the stages is performed using the markers CD27 and CD11b on gated NK cells. The significance of this finding is underpinn ...

Being exposed to food products, pathogens and harmless commensal bacteria, the mucosal immune system faces a constant challenge. Therefore, maintenance of a homeostatic balance is required to achieve tolerance to harmless bacteria and their products and to induce potent immunity to i ...

The term uterine natural killer (uNK) cell is applied in mice to an abundant but transient NK cell population that undergoes unique, terminal differentiation within embryo implantation sites during endometrial decidualization and pregnancy. In mice, decidualization is induced by ...

Natural killer (NK) cells account for 70% of the leukocytes in the mucosal lining of the uterus (the decidua) in the first trimester of pregnancy. They are CD56superbright granulated cells expressing a repertoire of Killer-cell Immunoglobulin-like Receptors (KIR) skewed towards reco ...

Frequently direct measurement of proteins or their phosphorylation in intact cells is not possible, for instance, when cells are too few, frozen, or subject to degradation. We have demonstrated that tumor cells pour their DNA, RNA, and protein content into circulation because of turnover and ...

Alemtuzumab (Campath), the humanized rat monoclonal antibody that targets the CD52 surface antigen, is currently used for treatment of patients with resistant chronic lymphocytic leukemia. Monitoring levels of the antibody in plasma/serum could provide insight into the optimal ...

Chromosome translocations resulting in fusion genes have been implicated in leukemogenesis. The paradigm involves the fusion of the genes encoding BCR and ABL, leading to a constitutively active tyrosine kinase. The detection of BCR-ABL has been limited to fluorescence in situ hybrid ...

Acquired and intrinsic multidrug resistance is the major reason for the failure of anticancer chemotherapy. The most important component of clinical multidrug resistance is mediated by P-glycoprotein (Pgp), an ABC transporter encoded by the MDR1 gene and expressed on the membrane of tu ...

Application of monoclonal antibodies (MAb) for therapeutic purpose may induce the formation of human antibodies directed against the immunogenic epitopes, which are presented on the therapeutic MAb. Formation of such human antibodies mostly is an undesired side effect, but in the case ...

About 40 yr ago, two groups of investigators identified a new class of immunoglobulins, IgE. By exchanging their results and reagents, they proved that the immunoglobulin responsible for immediate hypersensitivity was IgE. From that day forward the science of allergy was greatly advanc ...

Cell fusion protocols that were developed by Kohler and Milstein in the mid-1970s and aimed at producing and characterization of mouse monoclonal antibodies (MAbs) remain the gold standard of hybridoma development. Despite tremendous progress in using MAbs in multiple research, dia ...

Recombinant antibody technology is a rapidly evolving field that enables the study and improvement of antibody properties by means of genetic engineering. Moreover, the functional expression of antibody fragments in Escherichia coli has formed the basis for antibody library gene ...

As a source of recombinant antigen, soluble constant fragment (Fc) fusion proteins have become valuable reagents for immunotherapy and laboratory investigations. Additional applications for these reagents include flow cytometry, immunohistochemistry, and in vitro acti ...

Flow cytometry is an automated analysis of cells passing in the fluid suspension through a laser light beam, which react with monoclonal antibodies specific for a variety of cell surface antigens. A specimen of peripheral blood, bone marrow, or other cell suspension is incubated with fluores ...