Long-term in vitro culture of Plasmodium vivax isolates from Madagascar maintained in Saimiri boliviensis blood

Abstract

Background Plasmodium vivax is the most prevalent human malaria parasite and is likely to increase proportionally as malaria control efforts more rapidly impact the prevalence of Plasmodium falciparum. Despite the prominence of P. vivax as a major human pathogen, vivax malaria qualifies as a neglected and under-studied tropical disease. Significant challenges bringing P. vivax into the laboratory, particularly the capacity for long-term propagation of well-characterized strains, have limited the study of this parasite’s red blood cell (RBC) invasion mechanism, blood-stage development, gene expression, and genetic manipulation.Methods and resultsPatient isolates of P. vivax have been collected and cryopreserved in the rural community of Ampasimpotsy, located in the Tsiroanomandidy Health District of Madagascar. Periodic, monthly overland transport of these cryopreserved isolates to the country’s National Malaria Control Programme laboratory in Antananarivo preceded onward sample transfer to laboratories at Case Western Reserve University, USA. There, the P. vivax isolates have been cultured through propagation in the RBCs of Saimiri boliviensis. For the four patient isolates studied to-date, the median time interval between sample collection and in vitro culture has been 454 days (range 166–961 days). The median time in culture, continually documented by light microscopy, has been 159 days; isolate AMP2014.01 was continuously propagated for 233 days. Further studies show that the P. vivax parasites propagated in Saimiri RBCs retain their ability to invade human RBCs, and can be cryopreserved, thawed and successfully returned to productive in vitro culture.Conclusions/significanceLong-term culture of P. vivax is possible in the RBCs of Saimiri boliviensis. These studies provide an alternative to propagation of P. vivax in live animals that are becoming more restricted. In vitro culture of P. vivax in Saimiri RBCs provides an opening to stabilize patient isolates, which would serve as precious resources to apply new strategies for investigating the molecular and cellular biology of this important malaria parasite.Electronic supplementary materialThe online version of this article (10.1186/s12936-017-2090-7) contains supplementary material, which is available to authorized users.

Risk Factors for Malaria Infection in Central Madagascar: Insights from a Cross-Sectional Population Survey

“…8 Here, we present a study investigating the parasite reservoir and infection risk factors in an area of central Madagascar known to be co-endemic for both Plasmodium falciparum and Plasmodium vivax clinical malaria. 9,10 Four human Plasmodium parasite species have been detected in Madagascar, with P. falciparum being predominant. 6,11 Parasite species-specific routine case reporting is not yet established, although it is a stated ambition in the latest strategic plan, making it difficult, at present, to assess species-specific contributions to malaria morbidity and mortality.…”

“…Here, we revisit an area of Madagascar where P. vivax infections were previously described in Duffy-negative patients, and where clinical P. vivax malaria exists alongside P. falciparum. 9,10 In this context, the present study addresses two principal issues: 1) the proportions and characteristics of submicroscopic infections missed by conventional point-of-care diagnostic tests to provide a more complete description of the parasite reservoir, and 2) P. vivax infection risks according to Duffy genotype.…”

Plasmodium vivax Biology: Insights Provided by Genomics, Transcriptomics and Proteomics

“…The application of such tools to in vitro cell or animal experimental models may help us explore further some aspects of P. vivax biology and the pathophysiology of disease (Table 3 ). Specifically, the development, establishment and validation of in vitro cell culture and animal experimental models (as the published Mazier et al, 1984 ; Hollingdale et al, 1985 ; Sattabongkot et al, 2006 ; Cui et al, 2009 ; Mikolajczak et al, 2015 ; Mehlotra et al, 2017 reports) to support P. vivax growth would overcome the current number of parasites available for projects in all omics branches. Clearly, this would positively impact on our understanding of several molecular mechanisms taking place in the parasite during cell invasion, development, stage transition, host immune system evasion and environmental stress responses (Table 3 ).…”

Plasmodium vivax in vitro continuous culture: the spoke in the wheel

“… GlutaMAX did indeed improve parasite viability, growth and development compared to traditional l -glutamine [ 15 ] 2017 Infected patients (cryopreserved isolates) Saimiri boliviensis and human blood Long-term 233 Parasites could re-invade monkey and human erythrocytes. Dulbecco’s Modified Eagle Medium (DMEM) was effective for P. vivax culture [ 69 ] …”

“…The use of Dulbecco’s Modified Eagle Medium (DMEM) for P. vivax culture supplemented with l -glutamine, HEPES and hypoxanthine has been reported recently. Parasitaemia was maintained for 233 days and was ended because of bacterial contamination [ 69 ]. The fluctuation in parasitaemia using DMEM was similar to that observed when the parasite has been grown in McCoy’s5A medium [ 14 ], suggesting that these media (McCoy’s 5A and DMEM) are useful for culturing and maintaining parasite maturation and replication in vitro.…”