Molecular detection and quantification of Plasmodium falciparum-infected human hepatocytes in chimeric immune-deficient mice

Foquet, Lander; Hermsen, Cornelus C.; Gemert, Geert-Jan van; Libbrecht, Louis; Sauerwein, Robert W.; Meuleman, Philip; Leroux‐Roels, Geert

doi:10.1186/1475-2875-12-430

Cited by 19 publications

(26 citation statements)

References 40 publications

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“…An inoculum of 2.4 × 10 5 P. falciparum NF54 sporozoites and 2.4 × 10 5 ΔMAEBL F11 sporozoites freshly isolated from mosquito hemolymph was injected by intravenous tail injection into the same three humanized mice. Livers were obtained 6 days postinfection from CO 2 -euthanized mice, and individual lobes were cut as described (Foquet et al, 2013). Lobes were pooled and emulsified into a singlecell suspension and flash frozen in liquid nitrogen for subsequent gDNA extraction.…”

Section: Immunofluorescence Microscopymentioning

confidence: 99%

“…To quantify parasite load in the chimeric livers, gDNA was isolated from the single-cell liver suspensions, and Taqman probe-based quantitative PCRs were performed as previously described (Alcoser et al, 2011;Foquet et al, 2013;. To specifically differentiate NF54 from ΔMAEBL genomes from the same mouse samples, the following oligonucleotides were used: For NF54 genomes: MAEBL_F (5′-GAATGGCAAAAGGAAGGTGA-3′) and MAEBL_R (5′-GTGCCCTCCCTTCATAAACA-3′) that bind internal to the MAEBL gene in NF54 but do not amplify a product using ΔMAEBL parasites.…”

Section: Measuring Exoerythrocytic Development In Humanized Micementioning

confidence: 99%

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AMA1 and MAEBL are important forPlasmodium falciparumsporozoite infection of the liver

Yang

Lopaticki

O'Neill

et al. 2017

Cellular Microbiology

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The malaria sporozoite injected by a mosquito migrates to the liver by traversing host cells. The sporozoite also traverses hepatocytes before invading a terminal hepatocyte and developing into exoerythrocytic forms. Hepatocyte infection is critical for parasite development into merozoites that infect erythrocytes, and the sporozoite is thus an important target for antimalarial intervention. Here, we investigated two abundant sporozoite proteins of the most virulent malaria parasite Plasmodium falciparum and show that they play important roles during cell traversal and invasion of human hepatocytes. Incubation of P. falciparum sporozoites with R1 peptide, an inhibitor of apical merozoite antigen 1 (AMA1) that blocks merozoite invasion of erythrocytes, strongly reduced cell traversal activity. Consistent with its inhibitory effect on merozoites, R1 peptide also reduced sporozoite entry into human hepatocytes. The strong but incomplete inhibition prompted us to study the AMA-like protein, merozoite apical erythrocyte-binding ligand (MAEBL). MAEBL-deficient P. falciparum sporozoites were severely attenuated for cell traversal activity and hepatocyte entry in vitro and for liver infection in humanized chimeric liver mice. This study shows that AMA1 and MAEBL are important for P. falciparum sporozoites to perform typical functions necessary for infection of human hepatocytes. These two proteins therefore have important roles during infection at distinct points in the life cycle, including the blood, mosquito, and liver stages.

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Section: Immunofluorescence Microscopymentioning

confidence: 99%

Section: Measuring Exoerythrocytic Development In Humanized Micementioning

confidence: 99%

AMA1 and MAEBL are important forPlasmodium falciparumsporozoite infection of the liver

Yang

Lopaticki

O'Neill

et al. 2017

Cellular Microbiology

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“…Two heterozygous uPA +/− -SCID mice, not engrafted with human hepatocytes, served as controls and were also challenged with P. falciparum sporozoites. Livers were collected either at 24 hpi or 5 days post-infection for detection of P. falciparum 18S DNA by quantitative real-time PCR ( Foquet et al, 2013 ). Both mice infected with WT Pf and 1 of the 2 mice infected with PfΔ b9 Δ slarp were positive for Pf 18S DNA at 24 hr post-infection, demonstrating successful sporozoite infection in human hepatocytes ( Figure 3 ).…”

Section: Resultsmentioning

confidence: 99%

A genetically attenuated malaria vaccine candidate based on P. falciparum b9/slarp gene-deficient sporozoites

Schaijk

Ploemen

Annoura

et al. 2014

eLife

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A highly efficacious pre-erythrocytic stage vaccine would be an important tool for the control and elimination of malaria but is currently unavailable. High-level protection in humans can be achieved by experimental immunization with Plasmodium falciparum sporozoites attenuated by radiation or under anti-malarial drug coverage. Immunization with genetically attenuated parasites (GAP) would be an attractive alternative approach. In this study, we present data on safety and protective efficacy using sporozoites with deletions of two genes, that is the newly identified b9 and slarp, which govern independent and critical processes for successful liver-stage development. In the rodent malaria model, PbΔb9ΔslarpGAP was completely attenuated showing no breakthrough infections while efficiently inducing high-level protection. The human PfΔb9ΔslarpGAP generated without drug resistance markers were infective to human hepatocytes in vitro and to humanized mice engrafted with human hepatocytes in vivo but completely aborted development after infection. These findings support the clinical development of a PfΔb9ΔslarpSPZ vaccine.DOI: http://dx.doi.org/10.7554/eLife.03582.001

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“…39,40,43 Successful transplantation of hu-heps into mice is determined by quantification of human albumin (Alb) in the serum of the mice, and up to 90% of the mouse liver parenchyma can become occupied by hu-heps. 39,43 Humanized mice have already been used extensively to study other hepatotropic pathogens (HBV, HCV, HDV and Plasmodium falciparum), [35][36][37]39,44,45 to test the human-specific metabolism and toxicity of medicinal compounds 46,47 and for the evaluation of novel antiviral therapies. 48,49 Several models of humanized mice were reported to sustain HEV infection, such as humanized uPA +/+ -SCID, uPA +/+ -NOG and FRG mice.…”

Section: Human Liver Chimeric Mice (Humanized Mice)mentioning

confidence: 99%

Updates in Hepatitis E virus (HEV) field; lessons learned from human liver chimeric mice

Sayed

Meuleman

2019

Reviews in Medical Virology

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Summary Hepatitis E virus (HEV) is the most common cause of viral hepatitis globally, and it is an emerging pathogen in developed countries. In vivo studies of HEV have long been hindered due to the lack of an efficient small animal model. Recently, human liver chimeric mice were described as an elegant model to study chronic HEV infection. HEV infection was established in mice with humanized liver that were challenged with stool preparations containing HEV genotype (gt)1 and/or gt3. An increase in viral load and the level of HEV Ag in mouse samples were markers of active infection. Plasma‐derived HEV preparations were less infectious. The kinetics of HEV ORF2 Ag during HEV infection and its impact on HEV diagnosis were described in this model. In addition, the nature of HEV particles and HEV ORF2 Ag were characterized. Moreover, humanized mice were used to study the impact of HEV infection on the hepatic innate transcriptome and evaluation of anti‐HEV therapies. This review highlights recent advances in the HEV field gathered from well‐established experimental mouse models, with an emphasis on this model as a tool for elucidating the course of HEV infection, the study of the HEV life cycle, the interaction of the virus with the host, and the evaluation of new anti‐HEV therapies.

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Molecular detection and quantification of Plasmodium falciparum-infected human hepatocytes in chimeric immune-deficient mice

Cited by 19 publications

References 40 publications

AMA1 and MAEBL are important forPlasmodium falciparumsporozoite infection of the liver

AMA1 and MAEBL are important forPlasmodium falciparumsporozoite infection of the liver

A genetically attenuated malaria vaccine candidate based on P. falciparum b9/slarp gene-deficient sporozoites

Updates in Hepatitis E virus (HEV) field; lessons learned from human liver chimeric mice

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