疾病及处理

Drug discovery programs heavily rely on assays adequately monitoring the activity of the drug on the �parasite stage targeted. So far, assays used to screen molecules active against Plasmodium falciparum parasites have mostly been based on measuring growth inhibition of asexual blood ...

We describe two improved assays for in vitro and in vivo screening of inhibitors and chemicals for antimalarial activity against blood stages of the rodent malaria parasite, Plasmodium berghei. These assays are based on the determination of bioluminescence in small blood samples that is p ...

CD8 T cells are critical mediators of protection against Plasmodium liver-stage infection. Most studies of the CD8 T cell response to whole parasite Plasmodium vaccines address a single T cell epitope in BALB/c mice, and thus provide limited information. Here, we describe a surrogate activa ...

T-cell receptor transgenic mice are powerful tools to study T cell responses to malaria parasites. They allow for a population of antigen specific T cells to be monitored during developing responses to immunization or parasite infection; this makes them particularly useful to study fund ...

This chapter describes a protocol to assess activation of human NK cells following in vitro stimulation with malaria-infected red blood cells. Activation is assessed by flow cytometry, staining for cell surface expression of CD69 and accumulation of intracellular IFN-γ. Procedures ...

We describe simple and sensitive in vitro and in vivo assays to analyze Plasmodium liver stage development using transgenic P. berghei parasites (PbGFP-Luccon), which express the bioluminescent reporter protein, luciferase. In these assays, parasite development in hepatocytes ...

Exoerythrocytic Plasmodium parasites infect hepatocytes and develop to huge multinucleated schizonts inside a parasitophorous vacuole. Finally, thousands of merozoites are formed and released into the host cell cytoplasm by complete disintegration of the parasitophor ...

Intravital fluorescence microscopy is an invaluable tool to study a dynamic phenomenon through its direct observation in living organisms. This technique can combine qualitative and quantitative analysis and has been capital to address long-standing questions about Plasmod ...

There is an important role for in vitro assays to better understand the initial steps of malaria infection. In this section, we describe both microscopy-based and flow cytometry-based sporozoite invasion, migration and development assays with the rodent malaria parasites, Plasmodi ...

Sporozoites, the stage of Plasmodium infectious to vertebrates when injected in the skin by a mosquito vector, are highly motile cells. Their unusual form of gliding motility is essential for infectivity, allowing the parasite to travel through both the mosquito and mammalian hosts, inva ...

We describe a technology for imaging the sequestration of infected red blood cells (iRBC) of the rodent malaria parasite Plasmodium berghei both in the bodies of live mice and in dissected organs, using a transgenic parasite that expresses luciferase. Real-time imaging of sequestered iRBC ...

Fluorescence-based techniques have been used extensively in the malaria field to study the functional role of nuclear organization and gene positioning in blood stages of the human malaria parasite, Plasmodium falciparum. In this chapter, we present optimized protocols for bromou ...

Chromatin immunoprecipitation (ChIP) studies have been used extensively in recent years to study the functional role of histone marks, variant histones, and other chromatin factors in gene expression in the human malaria parasite, Plasmodium falciparum. In this chapter, we present a C ...

Due to the A/T-richness of the genome of Plasmodium falciparum, expressing P. falciparum proteins in heterologous expression systems is challenging. In addition, many P. falciparum proteins have high cysteine content and high molecular weight, which further complicates expressi ...

Atomic Force Microscopy (AFM) is a powerful tool for exploring the interaction between ligands and receptors, as well as their exact locations on the red cell surface. Here we discuss current and future applications for AFM based single-molecule force spectroscopy to study adhesion of Plas ...

The altered deformability of erythrocytes infected with Plasmodium falciparum is central in malaria �pathogenesis, as it influences the hemodynamic properties of the infected cell and its retention in the spleen. Exported parasite proteins, as well as the shape and volume of the paras ...

An increased level of cytosolic free calcium (Ca2+) is an essential second messenger for apical organelle discharge in Plasmodium falciparum merozoites. Here, we describe a method for isolation of viable and invasive P. falciparum merozoites. We also describe methods to measure cytoso ...

Cellular imaging has reemerged in recent years as a powerful approach to provide researchers with a direct measure of essential molecular events in a cell’s life, ranging in scale from broad morphological observations of whole cells to intricate single molecule imaging. When combined wi ...

Metabolomics is an increasingly common analytical approach for investigating metabolic networks of pathogenic organisms. This may be of particular use in the study of parasitic infections due to the intrinsic metabolic connection between the parasite and its host. In vitro cultures ...

The central role played by protein phosphorylation in the regulation of eukaryotic cellular processes calls for detailed investigations of this phenomenon in malaria parasites. Here, we describe protocols to measure the activity of protein kinases (using either recombinant pro ...