抗体抗原实验

The production of homogenous antibodies with a defined specificity is an optimal goal of antibody preparation. This goal was achieved by Kohler and Milstein with the development of the technology for hybridoma production in 1975 (1).

Determination of class and subclass of monoclonal antibodies (MAbs) is important because several immunochemical and functional properties of antibodies depend on the isotype. Knowledge of the light-chain isotype (κ for 95% or λ for 5% of mouse antibodies) has limited practical value. In c ...

Chemotaxis is the process by which leukocytes are directed to sites of inflammation under the influence of a concentration gradient of the soluble chemotactic molecules. Upon encountering a chemotactic molecule, responding leukocytes begin to migrate directionally from regio ...

Most human and murine leukocytes express multiple chemokine receptors with overlapping ligand specificity. Therefore the creation of stable cell lines expressing specific chemokine receptors greatly facilitates chemokine receptor characterization, particularly ...

Numerous and diverse biological functions are regulated by chemokines. In addition to the well characterized proinflammatory activities such as integrin activation, chemotaxis, lipid mediator biosynthesis, superoxide radical production, and granule enzyme release (r ...

Endothelial cell migration and proliferation play an important role in the growth and development of new blood vessels, in the process known as neovascularization or angiogenesis. In the normal developing embryo, new blood vessels are formed by the directed movement of endothelial cel ...

Knowledge of a receptor’ pharmacology provides information which is often essential to unraveling the biological function of that receptor. The chemokine system represents a striking example as most receptors in the family bind multiple chemokines, often with complex overlappi ...

The intracellular ionic composition is closely related to cellular activity. The key roles in controlling cell metabolism are played by calcium, hydrogen, sodium, and potassium concentrations. It is generally recognized that cells spend a great amount of their energy on stringent reg ...

Locomotion of leukocytes is a complex event requiring coordinated activity of cytoskeletal, membrane, and adhesion systems (1). Prior to the migration of cells into the inflamed tissue, they have to pass the endothelium. This process involves several types of adhesion molecule interac ...

Basophils and mast cells play an important role in allergic disorders. Both of these cell types produce and secrete many mediators that play an obligatory role in immediate hypersensitivity reactions. Some of the mediators are preformed and stored in granules. An example is histamine. Other ...

Monocytes, macrophages, neutrophils, and eosinophils are able to generate and release reactive oxygen species. The reactive oxygen species are generated by reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase which is activated by a number of different soluble ...

The term “PI 3-kinase” is now applied to a growing family of proteins that are able to convert phosphatidylinositol (PtdIns), PtdIns(4)P and PtdIns(4,5)P2 into PtdIns(3)P, PtdIns(3,4)P2 and PtdIns(3,4,5)P3 respectively, by phosphorylating the D-3 position of the inositol head groups of pho ...

Chemokines (e.g., IL-8) were originally identified as chemotactic proteins obtained from different natural sources. These were purified by the use of a bioassay followed by N-terminal sequencing and then molecular cloning giving the full sequence and making it possible to express the ch ...

One of the more intriguing features of the hemopoietic system is the ability of a single phenotypically homogeneous stem cell to give rise—through differentiation and development—to all the phenotypically diverse mature immuno-hemopoietic cell types (1). The stem cell therefore s ...

The interaction of a select group of chemokines with glycosaminoglycans has been demonstrated previously, both by affinity coelectrophoresis (1) and by cell-surface binding assays and immobilized binding heparin assays (2). The significance of the interaction of chemokines wi ...

Binding of chemokines to their receptors induces a variety of biological responses, ranging from early and rapid events such as receptor phosphorylation and calcium influx to prolonged responses such as cellular migration (1,2). Downmodulation of chemokine receptors from the cell s ...

Endocytosis regulates the cell-surface expression of many seven transmembrane G-protein coupled receptors (7TM-GPCRs) and has been implicated in both desensitization following agonist-induced activation and resensitization (1,2). The internalization mechanism for ...

Monoclonal antibodies (MAbs) produced through the technique of somatic cell fusion are fundamental tools in the exploration of chemokine biology. This chapter details the procedures that were used to generate a panel of MAbs directed against the human chemokine RANTES (1,2). The general ...

Human immunodeficiency virus (HIV) is a member of the retro virus family, classified under the lentivirus genus. Retroviruses are enveloped RNA viruses, which contain a core of capsid proteins, viral RNA, and enzymes. All infectious retroviral virions contain an enzyme, reverse transcr ...

Monoclonal antibodies (MAbs) to chemokine receptors are providing remarkable insight into the roles these receptors play in basal leukocyte trafficking for immune response and surveillance as well as cell specific recruitment to sites of inflammation. This latter role in particu ...