免疫学技术

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 ...

One of the main reasons for interest in chemokines is the ease with which their expression can be documented in physiological settings involving leukocyte trafficking or in diseases characterized by inflammatory cell infiltration. In the case of most ELR-containing CXC chemokines, t ...

As the chemokine family continues to grow in number, it is becoming increasingly important to determine the distinct function of each molecule. This is particularly important because of the apparent overlapping biological activity of some chemokines in vitro. One approach to determi ...

Precursors of most secreted and cell surface molecules carry signal sequences at their amino termini. The method coined as signal sequence trap (1–3) takes advantage of the presence of N-terminal signal sequences in most precursor forms of secretory proteins and transmembrane protein ...

Eosinophils are leukocytes that contain characteristic cationic proteins in their granules that bind the acidic dye eosin. In contrast to neutrophils, eosinophils are minority cells in the blood and are predominantly tissue-dwelling cells found at sites in contact with the environ ...

Chemokines are a large family of secreted proteins that regulate the trafficking and activation of leukocytes by binding to G-protein-coupled receptors. In vitro studies of cloned and native receptors have shown that most chemokines bind to more than one receptor, and most chemokine rece ...

Animal models of allergic lung inflammation have been developed in an attempt to better define the pathogenesis of asthma. Using these models, important mechanistic questions that are difficult to pursue through clinical investigation have been addressed. Classically guinea pi ...

The suitability of any animal model of allergic airway inflammation is assessed by its ability to mimic human asthma. Thus, the ideal model would exhibit early and late antigen-induced airway responses, reversible bronchial hyperresponsiveness, peribronchial eosinophilia, and ...

In all medical and biological research, the ultimate aim is to understand pathological processes and to apply this understanding in the alleviation of human disease. Although laboratory-based molecular biology techniques are playing an ever increasing role, it is still necessary to u ...

In humans rapidly progressive glomerulonephritis is characterized by glomerular inflammation and the formation of glomerular crescents, composed of an infiltrate of mononuclear inflammatory cells and proliferating parietal epithelial cells. As disease progresses the ...

Recent progress in the collection of cDNA sequences by random sequencing reveals numerous genes of biological interest. Although a putative function may be predicted from deduced amino acid sequence, a rigorous proof of identity of a gene will require a demonstration of the appropriate bi ...

Escherichia coli is one of the most powerful and versatile hosts for highlevel protein production. Its well-characterized genetics and biochemistry have led to the development of many different systems for heterologous protein expression (1,2). In this chapter, we focus on the use of the T7 e ...

Here we describe the expression of chemokines in the periplasmic space of Escherichia coli (1–3), which leads to correctly folded proteins with no N-terminal modifications, in the case of RANTES, and with a yield of several hundred micrograms from a 1-L culture. Other expression systems that are g ...

Solid phase peptide synthesis (SPPS) is an alternative to DNA expression for generating proteins, such as chemokines (1–1). DNA databases and cDNA cloning has resulted in an explosion in the number of new chemokines from 1995-1998. However, for studies of the protein, knowing the DNA sequence is o ...

The majority of chemokines are highly basic, small proteins, with a molecular mass of around 8–10 kDa. Although they do not necessarily have a high level of homology at the primary sequence level, which can be as low as 20%, although it can also be as high as 90%, the three-dimensional structure of all the chemoki ...

The chemokine superfamily is a large group of small proteins, which share a limited amount of primary sequence identity, although a highly conserved three-dimensional subunit structure. The original members of the superfamily, Interleukin-8 and MCP-1 were purified over 10 years ago. Fo ...

Detailed understanding of chemokine action ultimately requires molecular cloning of chemokine receptor genes and cDNAs. Both forward and reverse genetic methods have been used for this purpose (1). Forward methods, which involve primary purification of a protein based on a defined ph ...

The immune system responds to antigenic stimulation with a complex array of molecular events involving antigen-presenting cells, B-cells, T-cells, and phagocytes. Cytokines and their respective receptors are intimately involved in regulating such immune responses. Their piv ...