Long Term Cultivation of Plasmodium falciparum in Aotus trivirgatus Erythrocytes *

E, Peterson; Sg, Langreth; Kenney, Shannon C.

doi:10.4269/ajtmh.1984.33.331

Cited by 6 publications

(6 citation statements)

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“…Both the wild-type strain FCR-3 and the D3 clone parasites were cultured continously in Aotus erythrocytes for 1 month before infection of monkeys. The erythrocytes of all monkeys in these experiments supported the growth in vitro of both wild-type and D3 parasites under standard culture conditions (23).…”

Section: Resultsmentioning

confidence: 68%

“…Parasite cultivation. Parasites (FCR-3 wild type and clone D3) grown in human 0+ erythrocytes with A' serum by the Trager-Jensen (28) method were adapted to culture in Aotus monkey erythrocytes (23). The adapted parasites were maintained in vitro in Aotus erythrocytes for 1 month before monkeys were infected.…”

Section: Methodsmentioning

confidence: 99%

“…Because of reports by J. Barnwell et al (3,4), M. Hommel et al (9,10), and P. David et al (7) of changes in antigenicity upon passage of malaria parasites through splenectomized primates, we did not adapt the K-clone and K+ wild-type parasites in this way. Instead, the parasites were adapted in vitro by continuous culture in owl monkey erythrocytes by recently developed modifications of human cell culture methods (23).…”

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confidence: 99%

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Pathogenicity, stability, and immunogenicity of a knobless clone of Plasmodium falciparum in Colombian owl monkeys

Langreth¹,

E²

1985

Infect Immun

115

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The pathogenicity, immunogenicity, and morphological stability of a knobless clone of strain FCR-3 of the human malaria parasite Plasmodium falkiparum was investigated in Aotus monkeys. An early knob-bearing (K+), wild-type isolate of strain FCR-3 and the D3 knobless (K-) clone were adapted to Aotus monkey erythrocytes in continuous culture, establishing the parasites in Aotus cells without exposure to in vivo cellular or humoral immune responses. All monkeys, intact or splenectomized, which were infected with wild-type FCR-3 adapted to Aotus cells in vitro, developed virulent infections and had to be drug treated. The intact nonsplenectomized animals which received knobless D3 cloned parasites did not develop virulent infections even after multiple infections. The splenectomized monkeys which received the K-D3 clone had virulent infections. Late-stage wild-type K+ parasites sequestered in both intact and splenectomized monkeys, whereas late-stage D3 K-parasites did not sequester in the splenectomized animals. These results suggest that two elements affected the pathogenicity of the malaria parasites in these experiments. Knobs on K+-infected erythrocytes enabled these parasites to sequester, presumably by attachment to capillary endothelium. When present, the spleen eliminated circulating Klate-stage erythrocytes, presumably by selection on the basis of their nondeformability. Although clone D3 Kparasites are nonvirulent in intact monkeys, they induced some immunological protection against challenge with wild-type K+ parasites. The surface morphology of K-infected erythrocytes remains unaltered throughout these experiments, suggesting that loss of knobs is a stable condition. 760 on August 2, 2020 by guest http://iai.asm.org/ Downloaded from

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Section: Resultsmentioning

confidence: 68%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Pathogenicity, stability, and immunogenicity of a knobless clone of Plasmodium falciparum in Colombian owl monkeys

Langreth¹,

E²

1985

Infect Immun

115

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“…This parasite strain failed to develop in erythrocytes from rhesus monkeys ( Macaca mulatta ) and guinea pigs. Peterson et al [ 117 ] also successfully cultivated several strains of P. falciparum in the erythrocytes of Aotus trivirgatus . Attempts to propagate P. falciparum strains in vitro using Saimiri sciureus (squirrel monkey) erythrocytes were met with limited success, i.e., up to three weeks of continuous culture [ 118 ].…”

Section: Erythrocytesmentioning

confidence: 99%

Cultivation of Asexual Intraerythrocytic Stages of Plasmodium falciparum

Basco

2023

Pathogens

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Successfully developed in 1976, the continuous in vitro culture of Plasmodium falciparum has many applications in the field of malaria research. It has become an important experimental model that directly uses a human pathogen responsible for a high prevalence of morbidity and mortality in many parts of the world and is a major source of biological material for immunological, biochemical, molecular, and pharmacological studies. Until present, the basic techniques described by Trager and Jensen and Haynes et al. remain unchanged in many malaria research laboratories. Nonetheless, different factors, including culture media, buffers, serum substitutes and supplements, sources of erythrocytes, and conditions of incubation (especially oxygen concentration), have been modified by different investigators to adapt the original technique in their laboratories or enhance the in vitro growth of the parasites. The possible effects and benefits of these modifications for the continuous cultivation of asexual intraerythrocytic stages of P. falciparum, as well as future challenges in developing a serum-free cultivation system and axenic cultures, are discussed.

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“…Studies have demonstrated that Plasmodium species are incapable of de novo biosynthesis of fatty acids and cholesterol and must obtain it from the host [ 33 , 34 ]. It is also known that Aotus serum does not support in vitro culture of P. falciparum [ 35 – 37 ], possibly to low concentrations of essential nutrients such as cholesterol and lipids, that are known to be in balance both in serum and red blood cells [ 34 ]. Recently however, it was established that malaria parasites are in high demand for lipids during replication and enlargement of their parasitophorous vacuole membrane (PVM) in the liver [ 38 ], but little is known about the demands for lipids and cholesterol of P. vivax parasites during replication in ex vivo culture.…”

Section: Introductionmentioning

confidence: 99%

On the survival of 48 h Plasmodium vivax Aotus monkey-derived ex vivo cultures: the role of leucocytes filtration and chemically defined lipid concentrate media supplementation

Obaldía

Núñez

2020

Malar J

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Background Filtration of leukocytes (WBCs) is a standard practice of malaria ex vivo cultures. To date, few studies have considered the effect of filtration or the lack thereof on the survival of Plasmodium vivax ex vivo cultures through one cycle of maturation. This study investigates the effect of WBC filtration and culture media supplementation on the survival of 48–72 h ex vivo cultures. Methods Using parasitaemia density, the study compares the survival of Plasmodipur ® filtered, filter-retained or washed ex vivo cultures, maintained with McCoy’s5A medium supplemented with 25% serum alone or 20% in combination with 5% chemically defined lipid concentrate (CDLC), and in washed ex vivo cultures plus GlutaMAX™, benchmarked against IMDM™ or AIM-V™ media; also, assessed the survival of ex vivo cultures co-cultivated with human red blood cells (hRBCs). Results After 48 h of incubation a statistically significant difference was detected in the survival proportions of filtered and the filter-retained ex vivo cultures supplemented with serum plus CDLC ( p = 0.0255), but not with serum alone ( p = 0.1646). To corroborate these finding, parasitaemias of washed ex vivo cultures maintained with McCoy’s5A complete medium were benchmarked against IMDM™ or AIM-V™ media; again, a statistically significant difference was detected in the cultures supplemented with CDLC and GlutaMAX™ ( p = 0.03), but not when supplemented with either alone; revealing a pattern of McCoy’s5A medium supplementation for Aotus -derived P. vivax cultures as follows: serum < serum + GlutaMAX™ < serum + CDLC < serum + CDLC + GlutaMAX™; confirming a key role of CDLC in combination with GlutaMAX™ in the enhanced survival observed. Lastly, results showed that co-cultivation with malaria-naïve hRBCs improved the survival of ex vivo cultures. Conclusions This study demonstrates that WBC filtration is not essential for the survival of P. vivax ex vivo cultures. It also demonstrates that McCoy’s5A complete medium improves the survival of Aotus -derived P. vivax ex vivo cultures, with no significant difference in survival compared to IMDM and AIM-V media. Finally, the study demonstrates that co-cultivation with hRBCs enhances the survival of ex vivo cultures. These findings are expected to help optimize seeding material for long-term P. vivax in vitro culture.

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Long Term Cultivation of Plasmodium falciparum in Aotus trivirgatus Erythrocytes *

Cited by 6 publications

References 0 publications

Pathogenicity, stability, and immunogenicity of a knobless clone of Plasmodium falciparum in Colombian owl monkeys

Pathogenicity, stability, and immunogenicity of a knobless clone of Plasmodium falciparum in Colombian owl monkeys

Cultivation of Asexual Intraerythrocytic Stages of Plasmodium falciparum

On the survival of 48 h Plasmodium vivax Aotus monkey-derived ex vivo cultures: the role of leucocytes filtration and chemically defined lipid concentrate media supplementation

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