A transgenic Plasmodium falciparum NF54 strain that expresses GFP–luciferase throughout the parasite life cycle

Vaughan, Ashley M.; Mikolajczak, Sebastian A.; Camargo, Nelly; Lakshmanan, Viswanathan; Kennedy, Mark; Lindner, Scott E.; Miller, Jessica L.; Hume, Jen C. C.; Kappe, Stefan H. I.

doi:10.1016/j.molbiopara.2012.10.004

Cited by 62 publications

(85 citation statements)

References 19 publications

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“…No significant difference was observed in albumin levels or human GAPDH gene expression in the FRG huHep mice showing equal human hepatocyte repopulation between the mice. P. falciparum luciferase-expressing (NF54HT-GFP-luc) sporozoites (15) were introduced either by tail vein injection of 1 × 10 6 sporozoites or by the bite of 50 infected mosquitoes. Six days after challenge, when liver burden is highest, liver stage P. falciparum infection was measured using in vivo bioluminescence imaging (IVIS), as well as liver harvest followed by quantification of parasite 18S rRNA by quantitative reverse transcription PCR (qRT-PCR) ( Figure 1C).…”

Section: Resultsmentioning

confidence: 99%

“…The P. falciparum NF54HT-GFP-luc parasite, which constitutively expresses luciferase under the EF1a promoter (15), was maintained in RPMI 1640 (25 mM HEPES, 2 mM L-glutamine) supplemented with 50 μM hypoxanthine plus 10% human serum and subcultured in 5% O + human erythrocytes (Interstate Blood Bank). Mature sexual-stage gametocytes were induced as previously described (11).…”

Section: P Falciparum Sporozoite Production and Mouse Infectionmentioning

confidence: 99%

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Assessing drug efficacy against Plasmodium falciparum liver stages in vivo

et al. 2018

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Section: Resultsmentioning

confidence: 99%

Section: P Falciparum Sporozoite Production and Mouse Infectionmentioning

confidence: 99%

Assessing drug efficacy against Plasmodium falciparum liver stages in vivo

et al. 2018

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“…The infected mice were used to feed female Anopheles stephensi mosquitoes after gametocyte exflagellation was observed. P. falciparum GFP-luc parasites were maintained as previously described (34). Briefly, in vitro P. falciparum NF54HT-GFP-luc bloodstage cultures were maintained in RPMI 1640 (25 mM HEPES, 2 mM L-glutamine) supplemented with 50 M hypoxanthine and 10% Aϩ human serum in an atmosphere of 5% CO 2 , 5% O 2 , and 90% N 2 .…”

Section: Methodsmentioning

confidence: 99%

“…Finally, utilizing a recently developed, robust human liver-chimeric mouse model that supports infection with luciferase-expressing P. falciparum sporozoites (34,35), we demonstrate Abmediated inhibition of P. falciparum liver infection.…”

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confidence: 86%

Model forIn VivoAssessment of Humoral Protection against Malaria Sporozoite Challenge by Passive Transfer of Monoclonal Antibodies and Immune Serum

et al. 2014

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fEvidence from clinical trials of malaria vaccine candidates suggests that both cell-mediated and humoral immunity to pre-erythrocytic parasite stages can provide protection against infection. Novel pre-erythrocytic antibody (Ab) targets could be key to improving vaccine formulations, which are currently based on targeting antigens such as the circumsporozoite protein (CSP). However, methods to assess the effects of sporozoite-specific Abs on pre-erythrocytic infection in vivo remain underdeveloped. Here, we combined passive transfer of monoclonal Abs (MAbs) or immune serum with a luciferase-expressing Plasmodium yoelii sporozoite challenge to assess Ab-mediated inhibition of liver infection in mice. Passive transfer of a P. yoelii CSP MAb showed inhibition of liver infection when mice were challenged with sporozoites either intravenously or by infectious mosquito bite. However, inhibition was most potent for the mosquito bite challenge, leading to a more significant reduction of liver-stage burden and even a lack of progression to blood-stage parasitemia. This suggests that Abs provide effective protection against a natural infection. Inhibition of liver infection was also achieved by passive transfer of immune serum from whole-parasite-immunized mice. Furthermore, we demonstrated that passive transfer of a MAb against P. falciparum CSP inhibited liver-stage infection in a humanized mouse/P. falciparum challenge model. Together, these models constitute unique and sensitive in vivo methods to assess serum-transferable protection against Plasmodium sporozoite challenge.

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“…These approaches are mutually exclusive in the same animal, though, as qPCR analysis requires animal euthanasia and liver extraction prior to blood-stage infection -resulting in a doubling of animal numbers if both liver burden and time to patency are to be measured. Recently, measurement of liver-stage burden without animal sacrifice has become possible through live bioluminescent imaging techniques and the generation of parasites expressing the firefly luciferase gene [34,[66][67][68][69][70]. This approach strikes a balance between sensitivity of measuring liver-stage burden while keeping the animal alive to measure parasite progression to bloodstage patency, thus reducing animal use and also enabling recurrent challenge for investigation of duration of protection and immunological memory.…”

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confidence: 99%

An Expanding Toolkit for Preclinical Pre-Erythrocytic Malaria Vaccine Development: Bridging Traditional Mouse Malaria Models and Human Trials

2016

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Malaria remains a significant public health burden with 214 million new infections and over 400,000 deaths in 2015. Elucidating relevant Plasmodium parasite biology can lead to the identification of novel ways to control and ultimately eliminate the parasite within geographic areas. Particularly, the development of an effective vaccine that targets the clinically silent pre-erythrocytic stages of infection would significantly augment existing malaria elimination tools by preventing both the onset of blood-stage infection/disease as well as spread of the parasite through mosquito transmission. In this Perspective, we discuss the role of small animal models in pre-erythrocytic stage vaccine development, highlighting how human liver-chimeric and human immune system mice are emerging as valuable components of these efforts. Malaria continues to cause significant morbidity and mortality despite renewed and concerted efforts to eliminate the causative agents, parasites of the genus Plasmodium. These efforts have largely focused on control of the Anopheles mosquito vectors, and antimalarial drug treatment of symptomatic infected individuals. The 214 million new malaria infections and 438,000 deaths due to malaria in 2015 were disproportionately borne by low income countries where malaria is endemic, and declines in malaria infections since 2000 have been slowest in countries with low resource availability and high malaria burden [1]. In the fight against malaria, effective vaccines preventing both disease and transmission will be important tools to achieve WHO goals of a 90% reduction in disease burden by 2030 [1,2]. In humans, malaria is caused predominantly by the parasite species Plasmodium falciparum and Plasmodium vivax, with the remainder of infections caused by three additional parasite species, Plasmodium malariae, Plasmodium ovale and Plasmodium knowlesi [1].Malaria parasite transmission occurs when a Plasmodium-infected Anopheles mosquito bites and by probing the skin for a blood meal, deposits motile sporozoite stages into the host. Sporozoites pass through the dermis using active motility, enter a capillary and then rapidly home to the liver. Once in the liver sporozoites enter the parenchyma and infect hepatocytes, wherein they then transmogrify into liver stages, which grow, replicate and ultimately differentiate into tens of thousands of first-generation merozoites. Merozoites of human malaria parasites emerge from the liver into the blood stream 7-10 days after transmission, where they undergo cyclic erythrocytic infection and intraerythrocytic replication. This blood-stage infection is responsible for all mortality and clinical symptoms of malaria, as well as infection of a new mosquito vector by sexual stage parasites for onward transmission [3,4]. Stages prior to blood-stage infection (i.e., the sporozoite and liver stages) are collectively known as preerythrocytic (PE) stages of infection and vaccine candidates targeting these stages are collectively termed PE vaccines. By preventing progression to ...

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A transgenic Plasmodium falciparum NF54 strain that expresses GFP–luciferase throughout the parasite life cycle

Cited by 62 publications

References 19 publications

Assessing drug efficacy against Plasmodium falciparum liver stages in vivo

Assessing drug efficacy against Plasmodium falciparum liver stages in vivo

Model forIn VivoAssessment of Humoral Protection against Malaria Sporozoite Challenge by Passive Transfer of Monoclonal Antibodies and Immune Serum

An Expanding Toolkit for Preclinical Pre-Erythrocytic Malaria Vaccine Development: Bridging Traditional Mouse Malaria Models and Human Trials

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