Viability and Infectivity of Plasmodium vivax Gametocytes in Short-Term Culture

Ramos, Glenda Quaresma; Baía-da-Silva, Djane Clarys; Lacerda, Marcus Vinícius Guimarães; Monteiro, Wuelton Marcelo; Lopes, Stefanie Costa Pinto

doi:10.3389/fcimb.2021.676276

Cited by 6 publications

(6 citation statements)

References 33 publications

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“…11:e4045 [ 44 ] 2022 Ramos Viability and infectivity of Plasmodium vivax gametocytes in short-term culture Front Cell Infect Microbiol. 11:676276 [ 53 ] 2022 Dinko Generation of Plasmodium falciparum gametocytes in vitro with specific consideration for field isolates Method Mol Biol. 2470:121–32 [ 40 ] …”

Section: Resultsmentioning

confidence: 99%

“…As an alternative, ex vivo cultures of gametocytes have been set up by optimizing the enrichment method of clinical isolates and culture conditions. Gametocytes from P. vivax clinical isolates were successfully cultured for 48 h after being purified using a Percoll gradient, KCl Percoll gradient or MACS after leukodepletion [ 51 – 53 ]. The addition of KCl to the Percoll gradient was thought to minimize dehydration of cells applied to the gradient [ 52 ].…”

Section: Resultsmentioning

confidence: 99%

“…AB+ and O+ pooled human sera were the most reported blood type used, followed by A+. IMDM + 10% human AB+ serum was used for ex vivo P. vivax culture [ 52 , 53 ]. Culture in the presence of 10% human serum is commonly used, and the corresponding mature gametocytemia ranges from 0.1% to 3% with stress-based induction and is 4% with conditional gametocytogenesis induction.…”

Section: Resultsmentioning

confidence: 99%

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Protocols for Plasmodium gametocyte production in vitro: an integrative review and analysis

et al. 2022

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Background The production of Plasmodium gametocytes in vitro is a real challenge. Many protocols have been described, but few have resulted in the production of viable and infectious gametocytes in sufficient quantities to conduct research on—but not limited to—transmission-blocking drug and vaccine development. The aim of this review was to identify and discuss gametocyte production protocols that have been developed over the last two decades. Methods We analyzed the original gametocyte production protocols published from 2000 onwards based on a literature search and a thorough review. A systematic review was performed of relevant articles identified in the PubMed, Web of Sciences and ScienceDirect databases. Results A total 23 studies on the production of Plasmodium gametocytes were identified, 19 involving in vitro Plasmodium falciparum, one involving Plasmodium knowlesi and three involving ex vivo Plasmodium vivax. Of the in vitro studies, 90% used environmental stressors to trigger gametocytogenesis. Mature gametocytemia of up to 4% was reported. Conclusions Several biological parameters contribute to an optimal production in vitro of viable and infectious mature gametocytes. The knowledge gained from this systematic review on the molecular mechanisms involved in gametocytogenesis enables reproducible gametocyte protocols with transgenic parasite lines to be set up. This review highlights the need for additional gametocyte production protocols for Plasmodium species other than P. falciparum. Graphical abstract

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Protocols for Plasmodium gametocyte production in vitro: an integrative review and analysis

et al. 2022

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“…In order to determine whether MB possesses a gametocytocidal action, the SMFA methodology was performed using a parasite culture pre-exposed to this drug. Parasite maintenance was carried out as described previously ( Ramos et al., 2021 ). After leukocyte depletion, for each MB concentration (5, 10, and 20 μM) and control group (without drug), 400 µl of the pellet was cultured in 25 cm 2 bottles using IMDM supplemented with 20% inactive AB human serum to a final hematocrit of 2%.…”

Section: Methodsmentioning

confidence: 99%

The activity of methylene blue against asexual and sexual stages of Plasmodium vivax

Fabbri

Ramos

Baía-da-Silva

et al. 2023

Front. Cell. Infect. Microbiol.

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Methylene blue (MB) is an alternative for combating drug-resistant malaria parasites. Its transmission-blocking potential has been demonstrated in vivo in murine models, in vitro, and in clinical trials. MB shows high efficacy against Plasmodium vivax asexual stages; however, its efficacy in sexual stages is unknown. In this study, we evaluated the potential of MB against asexual and sexual forms of P. vivax isolated from the blood of patients residing in the Brazilian Amazon. An ex vivo schizont maturation assay, zygote to ookinete transformation assay, direct membrane feed assay (DMFA), and standard membrane feed assay (SMFA) using P. vivax gametocytes with MB exposure were performed. A cytotoxicity assay was also performed on freshly collected peripheral blood mononuclear cells (PBMCs) and the hepatocyte carcinoma cell line HepG2. MB inhibited the P. vivax schizont maturation and demonstrated an IC50 lower than that of chloroquine (control drug). In the sexual forms, the MB demonstrated a high level of inhibition in the transformation of the zygotes into ookinetes. In the DMFA, MB did not considerably affect the infection rate and showed low inhibition, but it demonstrated a slight decrease in the infection intensity in all tested concentrations. In contrast, in the SMFA, MB was able to completely block the transmission at the highest concentration (20 µM). MB demonstrated low cytotoxicity to fresh PBMCs but demonstrated higher cytotoxicity to the hepatocyte carcinoma cell line HepG2. These results show that MB may be a potential drug for vivax malaria treatment.

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“…Discovery of candidate vaccine proteins and testing them continues, predominantly to develop P. falciparum vaccines [ 279 , 282 , 284 , 290 , 295 , 300 , 307 , 308 ]. Nonetheless, P. vivax research has been progressing at a competitive pace, even prior to the genome sequencing era, greatly aided by the use of both Aotus and Saimiri monkey infections and short-term in vitro blood-stage cultures permitting studies on P. vivax from human infected blood samples [ 303 , 309 , 310 ]. Over the span of several decades, these animal models have been critical for obtaining P. vivax parasites for genetic and biological analyses (reviewed in [ 19 , 20 , 311 ]), especially in light of the absence of a reliable and robust long-term blood-stage culture system for P. vivax [ 312 – 314 ], as has been available for P. falciparum for decades [ 315 ].…”

Section: Twenty-first Century—turning Point In Malaria Researchmentioning

confidence: 99%

Systems biology of malaria explored with nonhuman primates

Galinski

2022

Malar J

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Abstract“The Primate Malarias” book has been a uniquely important resource for multiple generations of scientists, since its debut in 1971, and remains pertinent to the present day. Indeed, nonhuman primates (NHPs) have been instrumental for major breakthroughs in basic and pre-clinical research on malaria for over 50 years. Research involving NHPs have provided critical insights and data that have been essential for malaria research on many parasite species, drugs, vaccines, pathogenesis, and transmission, leading to improved clinical care and advancing research goals for malaria control, elimination, and eradication. Whilst most malaria scientists over the decades have been studying Plasmodium falciparum, with NHP infections, in clinical studies with humans, or using in vitro culture or rodent model systems, others have been dedicated to advancing research on Plasmodium vivax, as well as on phylogenetically related simian species, including Plasmodium cynomolgi, Plasmodium coatneyi, and Plasmodium knowlesi. In-depth study of these four phylogenetically related species over the years has spawned the design of NHP longitudinal infection strategies for gathering information about ongoing infections, which can be related to human infections. These Plasmodium-NHP infection model systems are reviewed here, with emphasis on modern systems biological approaches to studying longitudinal infections, pathogenesis, immunity, and vaccines. Recent discoveries capitalizing on NHP longitudinal infections include an advanced understanding of chronic infections, relapses, anaemia, and immune memory. With quickly emerging new technological advances, more in-depth research and mechanistic discoveries can be anticipated on these and additional critical topics, including hypnozoite biology, antigenic variation, gametocyte transmission, bone marrow dysfunction, and loss of uninfected RBCs. New strategies and insights published by the Malaria Host–Pathogen Interaction Center (MaHPIC) are recapped here along with a vision that stresses the importance of educating future experts well trained in utilizing NHP infection model systems for the pursuit of innovative, effective interventions against malaria.

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Viability and Infectivity of Plasmodium vivax Gametocytes in Short-Term Culture

Cited by 6 publications

References 33 publications

Protocols for Plasmodium gametocyte production in vitro: an integrative review and analysis

Protocols for Plasmodium gametocyte production in vitro: an integrative review and analysis

The activity of methylene blue against asexual and sexual stages of Plasmodium vivax

Systems biology of malaria explored with nonhuman primates

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