Simple and sensitive antimalarial drug screening in vitro and in vivo using transgenic luciferase expressing Plasmodium berghei parasites

Franke-Fayard, Blandine; Djokovic, Dusan; Dooren, Maaike W. van; Ramesar, Jai; Waters, Andrew P.; Falade, Mofolusho O.; Kranendonk, Michel; Martinelli, Axel; Cravo, Pedro Vitor Lemos; Janse, Chris J.

doi:10.1016/j.ijpara.2008.05.012

Cited by 70 publications

(72 citation statements)

References 19 publications

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“…The red fluorescent P. yoelii clone contains the RedStar cassette integrated at the d-ssu-rrna locus. We used the P. berghei clone 676cl1 (PbGFP-LUC SCH ) expressing a GFP-luciferase fusion gene via the EEF1α promoter (25). C57BL/6, Swiss and hairless SKH1 mice were purchased from Charles River Laboratories.…”

Section: Methodsmentioning

confidence: 99%

“…We next imaged parasite growth in the skin using bioluminescence. For this, we used sporozoites of the transgenic P. berghei clone 676cl1 (PbGFP-LUC SCH ) expressing a GFP-luciferase fusion gene via the EEF1α promoter (25), which constitutively produces the fusion protein throughout the parasite life cycle. Sporozoites (2 × 10 4 ) were inoculated in the ear skin of mice and luciferin injected immediately before real-time whole-body imaging of mice using the IVIS system (26).…”

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confidence: 99%

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Development of the malaria parasite in the skin of the mammalian host

Gueirard

Tavares²,

Thiberge³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

122

109

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The first step of Plasmodium development in vertebrates is the transformation of the sporozoite, the parasite stage injected by the mosquito in the skin, into merozoites, the stage that invades erythrocytes and initiates the disease. The current view is that, in mammals, this stage conversion occurs only inside hepatocytes. Here, we document the transformation of sporozoites of rodentinfecting Plasmodium into merozoites in the skin of mice. After mosquito bite, ∼50% of the parasites remain in the skin, and at 24 h ∼10% are developing in the epidermis and the dermis, as well as in the immunoprivileged hair follicles where they can survive for weeks. The parasite developmental pathway in skin cells, although frequently abortive, leads to the generation of merozoites that are infective to erythrocytes and are released via merosomes, as typically observed in the liver. Therefore, during malaria in rodents, the skin is not just the route to the liver but is also the final destination for many inoculated parasites, where they can differentiate into merozoites and possibly persist.intravital imaging | Plasmodium | schizogony M alarial infection starts with the inoculation of Plasmodium sporozoites by mosquitoes probing the vertebrate skin for blood. The highly motile sporozoites eventually invade host target cells where they differentiate and divide into numerous merozoites, the parasite form that invades erythrocytes and initiates the pathogenic phase of malarial infection. The host cell type in which sporozoites transform into merozoites, however, differs between Plasmodium species. In species that infect birds, sporozoites differentiate inside macrophages primarily in the skin but also in the spleen, liver, and bone marrow (1). In species that infect mammals, sporozoites are known to differentiate only inside hepatocytes in the liver (2-4).The first demonstration that sporozoites of mammal-infecting Plasmodium species develop inside hepatocytes was made in 1948 after i.v. inoculation of sporozoites of P. cynomologi into rhesus monkeys (2). In addition to reporting fully mature parasites inside hepatocytes, the authors also documented the persistence of immature and dormant forms of the parasite in the liver several months after the initial inoculation, which they proposed to be the cause of relapses (5), and were later called hypnozoites (6). Subsequent work indicated that sporozoites of species that infect humans (7) also undergo complete development inside hepatocytes.Since these early studies, P. berghei and the related P. yoelii species, which infect rodents, have been used as practical and safe models for studying the pre-erythrocytic phase of malaria. These parasites were shown to differentiate in the liver of laboratory rodents (8), and the P. berghei/rodent system was used to demonstrate that the majority of sporozoites were inoculated by mosquitoes in the skin rather than directly into the blood circulation (9), as traditionally assumed. More recently, the generation of fluorescent P. berghei parasites, alo...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Development of the malaria parasite in the skin of the mammalian host

Gueirard

Tavares²,

Thiberge³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

122

109

View full text Add to dashboard Cite

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“…Drug dose, expressed in mg/kg of body weight, was adjusted at the time of administration according to the weight of each mouse. The dose was based on the ED90 (5 mg/kg) on PbANKA infected mice [10].…”

Section: Standard Antimalarial Drugmentioning

confidence: 99%

Effect of Crude Leaf Extract of Bauhinia strychnifolia in BALB/c Mice Infected with Plasmodium berghei

Chutoam¹,

Klongthalay²,

Somsak³

2015

Malaria Contr Elimination

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Continuous emergence of antimalarial drug resistant malaria parasites and their rapid spread across the globe warrant urgent search for new antimalarials. Traditional medicinal plants have been the main sources for screening active phytochemicals against malaria. Accordingly, this study was aimed at evaluating the antimalarial activity of crude leaf extract of Bauhinia strychnifolia against Plasmodium berghei infected mice. Aqueous crude leaf extract of B. strychnifolia have been prepared and tested for acute toxicity and antimalarial efficacy in P. berghei ANKA infected BALB/c mice. At three oral doses of 100, 500 and 1,000 mg/kg of extract were safe, chemosuppressive and thus prevented body weight loss, packed cell volume reduction and increased mice mean survival time in a dosedependent manner compared to the untreated control group. The maximum efficacious extract was found at the dose of 1,000 mg/kg which prolonged mean mouse survival past day 26 of infection with all the mice in this group having the highest parasitemia suppression rate (85%). This study suggests that the crude leaf extract of this plant have promising antimalarial activity against P. berghei in a dose dependent manner, which supports the folkloric use of the plant for treating malaria.

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“…The introduction of a number of bioluminescence-based methods enables faster and less subjective quantification of the P. berghei burden when compared with traditional microscopy-based approaches. [123][124][125][126] These methods, however, do not permit the comparison of multiple mutants simultaneously. A flow cytometry-based in vivo competition assay has been developed that enables the analysis of blood-stage development of three differently coloured fluorescent parasite lines within a single mouse.…”

Section: Next Generationmentioning

confidence: 99%

Towards genome-wide experimental genetics in thein vivomalaria model parasitePlasmodium berghei

Matz

Kooij

2015

Pathogens and Global Health

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Plasmodium berghei was identified as a parasite of thicket rats (Grammomys dolichurus) and Anopheles dureni mosquitoes in African highland forests. Successful adaptation to a range of rodent and mosquito species established P. berghei as a malaria model parasite. The introduction of stable transfection technology, permitted classical reverse genetics strategies and thus systematic functional profiling of the gene repertoire. In the past 10 years following the publication of the P. berghei genome sequence, many new tools for experimental genetics approaches have been developed and existing ones have been improved. The infection of mice is the principal limitation towards a genome-wide repository of mutant parasite lines. In the past few years, there have been some promising and most welcome developments that allow rapid selection and isolation of recombinant parasites while simultaneously minimising animal usage. Here, we provide an overview of all the currently available tools and methods.

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Simple and sensitive antimalarial drug screening in vitro and in vivo using transgenic luciferase expressing Plasmodium berghei parasites

Cited by 70 publications

References 19 publications

Development of the malaria parasite in the skin of the mammalian host

Development of the malaria parasite in the skin of the mammalian host

Effect of Crude Leaf Extract of Bauhinia strychnifolia in BALB/c Mice Infected with Plasmodium berghei

Towards genome-wide experimental genetics in thein vivomalaria model parasitePlasmodium berghei

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