免疫学技术

In mammals, the genes encoding the variable (V) domains of the immunoglobulin heavy (H) chain are assembled during lymphocyte development by rearrangements of variable (VH), diversity (DH), and junctional (JH) gene segments (1). Selection of these gene elements from their corresponding ...

Enzyme-linked immunosorbent assay (ELISA, EIA) is a highly versatile and sensitive technique that can be used for quantitative as well as qualitative determination of almost any antigen or antibody. Reagents are stable, non-radioactive and, in most cases, commercially available. Ow ...

The measurement and manipulation of cytosolic free Ca2+ permits the investigation of the mechanisms of generation of the Ca2+ signal and cellular responses to these Ca2+ signals within living neutrophils. The optical methods most applicable to neutrophils, which will be discussed here, ...

The design of novel vaccines and strategies to combat infectious disease requires an understanding of the interactions between pathogen and host. Biological interactions in vivo often rely on specific recognition mechanisms that begin with a binding step. The development of biosen ...

The PorA or class 1 protein is one of the major meningococcal outermembrane proteins (OMPs). It is one of the two porins found in this organism, the other one being the PorB or class 2/3 protein. It folds into a 16-stranded β-barrel structure, which is now well-established for bacterial porins, in which seven ...

Lipopolysaccharide (LPS) is a major component of the meningococcal outer membrane. It consists of a hexa-acylated glucosamine disaccharide substituted at both ends with diphosphoethanolamine, to which an oligosaccharide chain of up to 10 sugar residues is attached (1,2). It lacks a lo ...

The outer membrane of Neisseria meningitidis contains a variety of proteins with the potential for inclusion in new meningococcal vaccines (1). Studies on the vaccine potential of these proteins would be facilitated by the production of pure recombinant protein, free from other compon ...

Proteins with intrinsic mitogenic properties are widely represented in prokaryotes, such as in different Streptococcus species (1–3), Candida albicans (4), and Eikenella corrodens (5). Specifically, several bacterial porins of Escherichia coli, Shigella dysenteriae, Salm ...

Although capsular polysaccharide-based vaccines are effective at reducing the incidence of meningococcal disease caused by serogroups A, C, Y, and W135 (1–3), immunization against serogroup B disease using similar strategies has proven unsuccessful (4,5). The primary reason for th ...

There are many in vitro systems for the study of meningococcal pathogenesis, but it is only in animal models of infection that the interactions of the bacteria with whole tissues and the humoral and cellular immune systems can be assessed. Animal-infection models are also of great importance for ...

In common with most infectious agents, meningococci invade mucosal membranes before causing disease (1). An attractive vaccine strategy is therefore to induce immunological responses at the mucosal surfaces, with the possibility of blocking microbial invasion, as well as to induce ...

The molecular analysis of cell signaling pathways requires an understanding of what each component in the pathway is, the manner in which they interact, and how these interactions serve to propagate the signal. Cytokine signaling begins with the binding of an extracellular cytokine to a spe ...

Among cytokine families, the tumor necrosis factor (TNF) superfamily stands out as the one that contains several members with cytotoxic activity (1–3). The best characterized is the prototypic member TNF-α, which is cytotoxic to a range of tumor cells, but not to most types of normal cells (1,4). How ...

It is well known that interferons (IFNs) have antiviral activity (1,2). In fact, the definition of interferon springs from this observation. Although it is reported that other cytokines, e.g., tumor necrosis factor (TNF) (3), may exert antiviral effects, biological assays based on antiviral a ...

The hematopoietic growth factor granulocyte colony-stimulating factor (G-CSF) has been in the clinic since 1986, when the clinical development of bacterially synthesized recombinant filgrastim began (1). A glycosylated form of G-CSF (lenograstim) has been approved for use in Euro ...

Leukocytes play an important role in inflammation and cancer. They are attracted to the place of injury by chemotactic factors, e.g., plasma complement factors C5a and C3a, and chemotactic cytokines or chemokines, which are produced locally at the site of infection. Chemokines are produced by a ...

Tumor necrosis factor-α (TNF-α) is one of the most pleiotropic of cytokines, acting as a host defense factor in myriad immunological and inflammatory responses and antitumor activity (1–3). The cytotoxic effects of TNF-α are primarily mediated through TNF-R1 and the receptor-associated ...

Cytokines, small soluble molecules, are secreted by many cells including cells of the immune system and the central nervous system (Figs. 1 and 2). Immunohistochemical detection of cytokines in tissues is hampered by the intrinsic properties of these molecules, i.e., their expression lev ...

Cytokines are a diverse group of proteins and glycoproteins that promote intercellular communication among leukocytes or between leukocyte and other cell types (1). Although most cytokines are soluble proteins, they can infrequently be found expressed at the cell surface. Cytokin ...

Methods for detection of cytokines and chemokines at a protein level in individual cells based on immunofluorescence or immunohistochemical (1–16) staining techniques have been developed in our laboratory. The difficulties in these procedures lie in the critical steps of fixation ...