实验动物技术

Subtractive hybridization techniques are designed to deplete one cDNA population (the “target”) of sequences common to a second group of cDNAs (the “driver”), thereby effectively enriching the target population for unique sequences. The applications of this powerful methodology ...

The discovery by Burke et al. in 1987 (1) that yeast can accept large pieces of heterologous DNA as yeast artificial chromosomes (YAC) marked a milestone in the analysis of complex genomes. While standard prokaryotic cloning systems, such as plasmids and cosmids, are limited by the size of the DNA frag ...

Genetic transformation of an organism is a fundamental investigational tool that allows the researcher to gain an insight into the basic cellular biology. Numerous areas of parasite biology can be addressed simply by introducing plasmids that are maintained as episomes into the cell of c ...

Gene targeting, the technology that permits inactivation or modification of a gene by homologous recombination, has been reproducibly established in two plasmodial species: Plasmodium falciparum (1), the most important pathogen for humans, and Plasmodium berghei (2), which infe ...

Vaccination is the most cost-effective measure to control the pathological effects of an infectious agent (1,2). With the combined use of molecular cloning and of modern sequencing methods, the sequence of many protein components present in different infectious agents have been eluci ...

There is increasing recognition that the unique antigenicities of the different stages of the Plasmodium spp. life cycle, the requirement for distinct immune mechanisms targeting these different stages, the immense allelic variation of parasites in the field, and the genetic hetero ...

To determine the immunogenicity of vaccine candidates, it is essential to establish an accurate and convenient methodology for assessing the level of antigen-specific T-cell responses, and several methodologies have been developed. These include the T-cell proliferative assay ...

In all rodent models of erythrocytic-stage malaria infections, the T-cell response to the parasite has been shown to be crucial for the development of protective immunity (1-5) and in some cases is thought to play a role in the pathology of malaria (6,7). To determine which effector functions of T cells m ...

CD4+helper T cells play a central role in the development of malaria immune responses and a large number of epitopes from the sporozoite, sexual, and asexual stage of malaria proteins have been tabulated in both mice and humans (1-8). It is clear that the incorporation of antigenic determinants sti ...

CD4+T cells play an important role in immunity to blood-stage malaria parasites and in disease pathogenesis. The role of CD4+T cells has been demonstrated by selective depletion in vitro (1-3), by adoptive transfer of T cells to immunodeficient mice (1,4-7), and by the ability of human T cells to inhib ...

There is evidence in rodent models of malaria and indirect evidence in human malaria of the role of T cells in protective immunity to the disease (13) and in the pathology of malaria infections (45)). Measurement of this T-cell response is important for two reasons: to determine which parasite protei ...

The advent of molecular techniques for the first time allowed for the specific biological characterization and a more clear understanding of the function of molecules synthesized by leukocytes termed cytokines. In addition, availability of the recombinant forms of these cytokines ...

Mosquito injection of Plasmodium sporozoites into the bloodstream is followed by a short transit period before invasion of the hepatocyte, the only host cell expressing class I major histocompatibility complex (MHC) molecules. Liver stages develop within the hepatocyte for perio ...

Townsend and colleagues (1) were the first to demonstrate, in 1984, that target cells that had been transfected with single viral RNA segments could be specifically recognized by cytotoxic T lymphocytes (CTL). Recombinant vaccinia viruses that expressed single-gene products were sub ...

Nitric oxide (NO) and the related species nitrosothiols, have multiple important physiological and pathological roles in health and disease that have been extensively reviewed elsewhere (1-6). In brief, NO is involved in modulating or mediating host resistance to tumors and microbes, ...

Our results have emphasized the potential involvement of the antibody-dependent cellular inhibition (ADCI) mechanism in the protective effect of antibodies mediating acquired immunity to Plasmodium falciparum malaria (1). The ADCI mechanism consists of a cooperation betwe ...

As described in Chapter 42, niric oxide (NO) is synthesized from the amino acid l-arginine by the actions of a family of enymes, the NO synthases (NOS), each isoform of which is encoded by a separate gene. Two NOS isoforms are calcium-dependent and constitutively expressed and produce low levels of NO: NO ...

Because it has been 25 yr since the successful cultivation of Plasmodium falciparum (1), most researchers do not remember how difficult it was to work with malaria parasites, especially in vitro. Before the development of current methods, malaria parasite cultures were always short-term, ...

Lipopolysaccharides (LPS) are a complex class of glycolipids that can trigger a cascade of immunological responses in mammals, including endotoxic effects and serum antibody production (1). LPS have been found to exhibit a common molecular architecture consisting of at least two dist ...

With the advent of new techniques to explore bacterial pathogenesis, such as signature tagged mutagenesis and site-directed mutagenesis coupled with the whole genome sequence and bioinformatics, there is a need for clear descriptions of the available mutagenesis tools for indivi ...