Improving in vitro continuous cultivation of Plasmodium cynomolgi, a model for P. vivax

Christensen, Peter; Annie, Racklyeft; Ward, Kurt E.; Matheson, Jessica; Rossarin, Suwanarusk; Adeline, Chua C.Y.; Kaneko, O.; Lin, Aung Htin; Laurent, Rénia; Amanzougaghene, Nadia; Victor, Magneron; Julien, Lemaitre; Roger, Le Grand; Dennis, Kyle; Pablo, Bifani; Gregory, M. Cook; Georges, Snounou; Russell, Bruce

doi:10.1016/j.parint.2022.102589

Cited by 9 publications

(6 citation statements)

References 12 publications

(10 reference statements)

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“…Nevertheless, genome-scale mutagenesis screen in P. falciparum identified Pf MCA3 gene as essential for the asexual blood-stage [ 30 ] and, more recently, a marked involvement of Pb MCA2 in the sexual stage development was shown using P. berghei knockout parasites [ 13 ], supporting a pivotal participation of metacaspases in different phases of Plasmodium life cycle. In this context, studies focusing the metacaspases of the simian malaria parasite P. cynomolgi , which has been proposed as a model system for in vivo and in vitro research on P. vivax [ 31 , 32 ], could bring some light into the functionality of Plasmodium metacaspases, especially the Pv MCAs.…”

Section: Resultsmentioning

confidence: 99%

Profile of metacaspase gene expression in Plasmodium vivax field isolates from the Brazilian Amazon

Blanco,

de Souza,

Martins

et al. 2024

Mol Biol Rep

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Background Metacaspases comprise a family of cysteine proteases implicated in both cell death and cell differentiation of protists that has been considered a potential drug target for protozoan parasites. However, the biology of metacaspases in Plasmodium vivax − the second most prevalent and most widespread human malaria parasite worldwide, whose occurrence of chemoresistance has been reported in many endemic countries, remains largely unexplored. Therefore, the present study aimed to address, for the first time, the expression pattern of metacaspases in P. vivax parasites. Methods and results P. vivax blood-stage parasites were obtained from malaria patients in the Brazilian Amazon and the expression of the three putative P. vivax metacaspases (PvMCA1-3) was detected in all isolates by quantitative PCR assay. Of note, the expression levels of each PvMCA varied noticeably across isolates, which presented different frequencies of parasite forms, supporting that PvMCAs may be expressed in a stage-specific manner as previously shown in P. falciparum. Conclusion The detection of metacaspases in P. vivax blood-stage parasites reported herein, allows the inclusion of these proteases as a potential candidate drug target for vivax malaria, while further investigations are still required to evaluate the activity, role and essentiality of metacaspases in P. vivax biology.

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Section: Resultsmentioning

confidence: 99%

Profile of metacaspase gene expression in Plasmodium vivax field isolates from the Brazilian Amazon

Blanco,

de Souza,

Martins

et al. 2024

Mol Biol Rep

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“…This low number of reports may have three possible reasons: (1) the characteristic of the inclusion criteria (as we excluded studies in invertebrate hosts, such as Plasmodium spp. Vectors); (2) difficulties in maintaining this parasite in culture, which often restricted clinical sampling in experiments [ 132 ]; and (3) the infected cell type of the host cell. The mature erythrocyte is one of the targets of Plasmodium spp.…”

Section: Resultsmentioning

confidence: 99%

A Systematic Review of Apicomplexa Looking into Epigenetic Pathways and the Opportunity for Novel Therapies

Brandão

Molento

2023

Pathogens

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Interest in host epigenetic changes during apicomplexan infections increased in the last decade, mainly due to the emergence of new therapies directed to these alterations. This review aims to carry out a bibliometric analysis of the publications related to host epigenetic changes during apicomplexan infections and to summarize the main studied pathways in this context, pointing out those that represent putative drug targets. We used four databases for the article search. After screening, 116 studies were included. The bibliometric analysis revealed that the USA and China had the highest number of relevant publications. The evaluation of the selected studies revealed that Toxoplasma gondii was considered in most of the studies, non-coding RNA was the most frequently reported epigenetic event, and host defense was the most explored pathway. These findings were reinforced by an analysis of the co-occurrence of keywords. Even though we present putative targets for repurposing epidrugs and ncRNA-based drugs in apicomplexan infections, we understand that more detailed knowledge of the hosts’ epigenetic pathways is still needed before establishing a definitive drug target.

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“…DBS were concurrently made using 20µL whole blood spotted on Whatman 3M filter paper and then stored in sealed bags with desiccant. A single donated P. cynomolgi -infected sample (20) obtained from a macaque host was frozen in glycerolyte and stored in liquid nitrogen prior to thawing, counting and immediately placing in DNA/RNA Shield TM .…”

Section: Methodsmentioning

confidence: 99%

Improved limit of detection for zoonoticPlasmodium knowlesiandP. cynomolgisurveillance using reverse transcription for total nucleic acid preserved samples or dried blood spots

Braima,

Piera,

Lubis

et al. 2024

Preprint

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Background:ZoonoticP. knowlesiandP. cynomolgisymptomatic and asymptomatic infections occur across endemic areas of Southeast Asia. Most infections are low-parasitemia, with an unknown proportion below routine microscopy detection thresholds. Molecular surveillance tools optimizing the limit of detection (LOD) would allow more accurate estimates of zoonotic malaria prevalence.Methods:An established ultra-sensitivePlasmodiumgenus quantitative-PCR (qPCR) assay targeting the 18S rRNA gene underwent LOD evaluation with and without reverse transcription (RT) forP. knowlesi, P. cynomolgiandP. vivaxusing total nucleic acid preserved (DNA/RNA ShieldTM) isolates and archived dried blood spots (DBS). LODs for selectedP. knowlesi-specific assays, and referenceP. vivax- andP. cynomolgi-specific assays were determined with RT. Assay specificities were assessed using clinical malaria samples and malaria-negative controls.Results:The use of reverse transcription improvedPlasmodiumspecies detection by up to 10,000-fold (Plasmodiumgenus), 2759-fold (P. knowlesi), 1000-fold (P. vivax) and 10-fold (P. cynomolgi). The median LOD with RT for the Kamau et al.Plasmodiumgenus RT-qPCR assay was ≤0.0002 parasites/µL forP. knowlesiand 0.002 parasites/µL for bothP. cynomolgiandP. vivax. The LODs with RT forP. knowlesi-specific PCRs were: Imwong et al. 18S rRNA (0.0007 parasites/µL); Divis et al. real-time 18S rRNA (0.0002 parasites/µL); Lubis et al. hemi-nested SICAvar (1.1 parasites/µL) and Lee et al. nested 18S rRNA (11 parasites/µL). The LOD forP. vivax- andP. cynomolgi-specific assays with RT were 0.02 and 0.20 parasites/µL respectively. For DBSP. knowlesisamples the median LOD for thePlasmodiumgenus qPCR with RT was 0.08, and without RT was 19.89 parasites/uL (249-fold change); no LOD improvement was demonstrated in DBS archived beyond 6 years. ThePlasmodiumgenus andP. knowlesi-assays were 100% specific forPlasmodiumspecies andP. knowlesidetection, respectively, from 190 clinical infections and 48 healthy controls. ReferenceP. vivax-specific primers demonstrated known cross-reactivity withP. cynomolgi.Conclusion:Our findings support the use of an 18S rRNAPlasmodiumgenus qPCR and species-specific nested PCR protocol with RT for highly-sensitive surveillance of zoonotic and humanPlasmodiumspecies infections.

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Improving in vitro continuous cultivation of Plasmodium cynomolgi, a model for P. vivax

Cited by 9 publications

References 12 publications

Profile of metacaspase gene expression in Plasmodium vivax field isolates from the Brazilian Amazon

Profile of metacaspase gene expression in Plasmodium vivax field isolates from the Brazilian Amazon

A Systematic Review of Apicomplexa Looking into Epigenetic Pathways and the Opportunity for Novel Therapies

Improved limit of detection for zoonoticPlasmodium knowlesiandP. cynomolgisurveillance using reverse transcription for total nucleic acid preserved samples or dried blood spots

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