Cryopreserved Plasmodium vivax and cord blood reticulocytes can be used for invasion and short term culture

Borlon, Céline; Russell, Bruce; Sriprawat, Kanlaya; Suwanarusk, Rossarin; Erhart, Annette; Rénia, Laurent; Nosten, François; D’Alessandro, Umberto

doi:10.1016/j.ijpara.2011.10.011

Cited by 46 publications

(61 citation statements)

References 25 publications

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“…22,35 Prior to cryopreservation, vivax malaria samples with parasitemia lower than 0.1% were subjected to 75% Percoll density gradient centrifugation to concentrate the parasitized erythrocytes. The cryopreserved samples were then thawed using the sodium chloride method.…”

Section: Rosetting Assay On Cryopreserved Samplesmentioning

confidence: 99%

“…[11][12][13][14][15][16][17] A renewed interest in vivax malaria and a better appreciation of its importance to public health has led to an increased number of studies examining particular aspects of P vivax pathogenesis. [18][19][20][21][22][23][24][25] Certainly in the case of P vivax, apart from some initial descriptions on P vivax rosetting 8,10 and its association with anemia, 26 little has been done to investigate the importance of rosetting to the survival of P vivax within the human host and the molecular mechanisms associated with the formation of rosettes in this species.…”

Section: Introductionmentioning

confidence: 99%

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Glycophorin C (CD236R) mediates vivax malaria parasite rosetting to normocytes

Lee

Malleret

Lau

et al. 2014

Blood

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Key Points• P vivax infected cells rosette exclusively to normocytes. Thus, rosetting does not directly facilitate P vivax merozoite invasion.• Glycophorin C (CD236R) mediates vivax malaria parasite rosetting. This finding will help in the search for the P vivax rosette ligand.Rosetting phenomenon has been linked to malaria pathogenesis. Although rosetting occurs in all causes of human malaria, most data on this subject has been derived from Plasmodium falciparum. Here, we investigate the function and factors affecting rosette formation in Plasmodium vivax. To achieve this, we used a range of novel ex vivo protocols to study fresh and cryopreserved P vivax (n 5 135) and P falciparum (n 5 77) isolates from Thailand. Rosetting is more common in vivax than falciparum malaria, both in terms of incidence in patient samples and percentage of infected erythrocytes forming rosettes. Rosetting to P vivax asexual and sexual stages was evident 20 hours postreticulocyte invasion, reaching a plateau after 30 hours. Host ABO blood group, reticulocyte count, and parasitemia were not correlated with P vivax rosetting. Importantly, mature erythrocytes (normocytes), rather than reticulocytes, preferentially form rosetting complexes, indicating that this process is unlikely to directly facilitate merozoite invasion. Although antibodies against host erythrocyte receptors CD235a and CD35 had no effect, Ag-binding fragment against the BRIC 4 region of CD236R significantly inhibited rosette formation. Rosetting assays using CD236R knockdown normocytes derived from hematopoietic stem cells further supports the role of glycophorin C as a receptor in P vivax rosette formation. (Blood. 2014;123(18):e100-e109)

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Section: Rosetting Assay On Cryopreserved Samplesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Glycophorin C (CD236R) mediates vivax malaria parasite rosetting to normocytes

Lee

Malleret

Lau

et al. 2014

Blood

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“…16 Indeed, the use of fresh and frozen fetal reticulocytes isolated from cord blood has allowed the development of reliable P vivax invasion assays. [17][18][19] To define the fine scale tropism of P vivax, we conducted ex vivo invasion assays on highly purified reticulocyte subsets (defined by TO staining and CD71 expression) isolated from cord blood. We further documented the remodeling of the reticulocyte following the invasion by the parasite.…”

Section: Introductionmentioning

confidence: 99%

Plasmodium vivax: restricted tropism and rapid remodeling of CD71-positive reticulocytes

Malleret

Li²,

Zhang

et al. 2015

Blood

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172

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Key Points• Plasmodium vivax merozoites preferentially infect a subgroup of reticulocytes generally restricted to the bone marrow.• Accelerated "maturation" of infected reticulocytes.Plasmodium vivax merozoites only invade reticulocytes, a minor though heterogeneous population of red blood cell precursors that can be graded by levels of transferrin receptor (CD71) expression. The development of a protocol that allows sorting reticulocytes into defined developmental stages and a robust ex vivo P vivax invasion assay has made it possible for the first time to investigate the fine-scale invasion preference of P vivax merozoites. Surprisingly, it was the immature reticulocytes (CD71 1) that are generally restricted to the bone marrow that were preferentially invaded, whereas older reticulocytes (CD71 2 ), principally found in the peripheral blood, were rarely invaded. Invasion assays based on the CD71 1 reticulocyte fraction revealed substantial postinvasion modification. Thus, 3 to 6 hours after invasion, the initially biomechanically rigid CD71 1 reticulocytes convert into a highly deformable CD71 2 infected red blood cell devoid of host reticular matter, a process that normally spans 24 hours for uninfected reticulocytes. Concurrent with these changes, clathrin pits disappear by 3 hours postinvasion, replaced by distinctive caveolae nanostructures. These 2 hitherto unsuspected features of P vivax invasion, a narrow preference for immature reticulocytes and a rapid remodeling of the host cell, provide important insights pertinent to the pathobiology of the P vivax infection. (Blood. 2015;125(8):1314-1324

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“…One of the obstacles to this system is that P. vivax mainly infects reticulocytes, which constitute only a small percentage of the red blood cells in adult donor blood [1]. Attempts to culture the parasites in vitro have therefore included provision of reticulocytes from Aotus monkeys [2], from human hemochromatosis patients [3] and human cord blood [4, 5, 6]. However, it is difficult to maintain these cultures for more than a few days and the sources for reticulocytes are not widely available.…”

Section: Introductionmentioning

confidence: 99%

Reticulocytes from cryopreserved erythroblasts support Plasmodium vivax infection in vitro

Furuya

Sá

Chitnis

et al. 2014

Parasitology International

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Plasmodium vivax is the most widely distributed human malaria parasite. Despite its importance, both clinical and basic research has been hampered by lack of a convenient in vitro culture system, in part due to the parasite’s infection preference of reticulocytes rather than mature erythrocytes. The use of reticulocyte-producing hematopoietic stem cell culture has been proposed for the maintenance of the parasite, but good numbers of reticulocytes and P. vivax parasites sufficient for practical use in research have been difficult to produce from this system. Here, we report an improved method of hematopoietic stem cell culture for P. vivax infection, which requires less time and produces higher or equivalent percentage of reticulocytes than previously reported systems. Reticulocytes were cultured from cryopreserved erythroblasts that were frozen after 8 days-cultivation of purified CD34+ cells from human umbilical cord blood. This method of production allowed the recovery of reticulocytes in a shorter time than with continuous stem cell culture. We obtained a relatively high percentage of peak reticulocyte production by using co-cultivation with a mouse stromal cell line. Using P. vivax mature stage parasites obtained from infected Aotus monkeys, we observed substantial numbers (up to 0.8 % of the total number of the cells) of newly invaded reticulocytes 24 hours after initial cultivation. The addition of fresh reticulocytes after 48 hours culture, however, did not result in significant increase of second cycle reticulocyte invasion. Assays of invasion inhibition with specific antibodies were successful with this system, demonstrating potential for study of biological processes as well as the conditions necessary for long-term maintenance of P. vivax in vitro.

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Cryopreserved Plasmodium vivax and cord blood reticulocytes can be used for invasion and short term culture

Cited by 46 publications

References 25 publications

Glycophorin C (CD236R) mediates vivax malaria parasite rosetting to normocytes

Glycophorin C (CD236R) mediates vivax malaria parasite rosetting to normocytes

Plasmodium vivax: restricted tropism and rapid remodeling of CD71-positive reticulocytes

Reticulocytes from cryopreserved erythroblasts support Plasmodium vivax infection in vitro

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