Red blood cells derived from peripheral blood and bone marrow CD34+ human haematopoietic stem cells are permissive to Plasmodium parasites infection

Fernández-Becerra, Carmen; Lelièvre, Joël; Ferrer, M. Carmen Olivé; Anton, Nuria; Thomson, Richard B.; Peligero, Cristina; Almela, María Jesús; Lacerda, Marcus; Herreros, Esperanza; Portillo, Hernando A. del

doi:10.1590/0074-0276108062013019

Cited by 15 publications

(18 citation statements)

References 11 publications

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“…JkRBCs Support Invasion by P. falciparum. We next tested the ability of jkRBCs to support P. falciparum invasion, as has been observed for other nucleated erythroid precursors (14,30,58). Indeed, we observed invasion into jkRBCs by two different strains of P. falciparum: the sialic acid-independent strain 3D7 (59) and the sialic acid-dependent strain Dd2 (Fig.…”

Section: Inhibition With Lsd1 Inhibitors Maintains An Undifferentiatementioning

confidence: 76%

CRISPR/Cas9 knockouts reveal genetic interaction between strain-transcendent erythrocyte determinants of Plasmodium falciparum invasion

Kanjee

Grüring

Chaand

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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During malaria blood-stage infections, parasites interact with the RBC surface to enable invasion followed by intracellular proliferation. Critical factors involved in invasion have been identified using biochemical and genetic approaches including specific knockdowns of genes of interest from primary CD34 hematopoietic stem cells (cRBCs). Here we report the development of a robust in vitro culture system to produce RBCs that allow the generation of gene knockouts via CRISPR/Cas9 using the immortal JK-1 erythroleukemia line. JK-1 cells spontaneously differentiate, generating cells at different stages of erythropoiesis, including terminally differentiated nucleated RBCs that we term "jkRBCs." A screen of small-molecule epigenetic regulators identified several bromodomain-specific inhibitors that promote differentiation and enable production of synchronous populations of jkRBCs. Global surface proteomic profiling revealed that jkRBCs express all known host receptors in a similar fashion to cRBCs and that multiple strains invade jkRBCs at comparable levels to cRBCs and RBCs. Using CRISPR/Cas9, we deleted two host factors, basigin (BSG) and CD44, for which no natural nulls exist. BSG interacts with the parasite ligand Rh5, a prominent vaccine candidate. A BSG knockout was completely refractory to parasite invasion in a strain-transcendent manner, confirming the essential role for BSG during invasion. CD44 was recently identified in an RNAi screen of blood group genes as a host factor for invasion, and we show that knockout results in strain-transcendent reduction in invasion. Furthermore, we demonstrate a functional interaction between these two determinants in mediating erythrocyte invasion.

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Section: Inhibition With Lsd1 Inhibitors Maintains An Undifferentiatementioning

confidence: 76%

CRISPR/Cas9 knockouts reveal genetic interaction between strain-transcendent erythrocyte determinants of Plasmodium falciparum invasion

Kanjee

Grüring

Chaand

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…In addition, it is possible to culture hematopoietic stem/progenitor cells (HSPC)/CD34 + cells to induce erythroid differentiation and consequently produce reticulocytes in vitro [8] . Reticulocytes generated from CD34 + cells from both bone marrow (BM) and peripheral blood mononuclear cells (PBMC) have been previously used for culturing P. vivax ; but the authors did not provide data regarding the efficiency of invasion and the development of the parasites in vitro [9] .…”

Section: Introductionmentioning

confidence: 99%

Hematopoietic Stem/Progenitor Cell Sources to Generate Reticulocytes for Plasmodium vivax Culture

Noulin

Manesia²,

Rosanas-Urgell

et al. 2014

PLoS ONE

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The predilection of Plasmodium vivax (P. vivax) for reticulocytes is a major obstacle for its establishment in a long-term culture system, as this requires a continuous supply of large quantities of reticulocytes, representing only 1–2% of circulating red blood cells. We here compared the production of reticulocytes using an established in vitro culture system from three different sources of hematopoietic stem/progenitor cells (HSPC), i.e. umbilical cord blood (UCB), bone marrow (BM) and adult peripheral blood (PB). Compared to CD34+-enriched populations of PB and BM, CD34+-enriched populations of UCB produced the highest amount of reticulocytes that could be invaded by P. vivax. In addition, when CD34+-enriched cells were first expanded, a further extensive increase in reticulocytes was seen for UCB, to a lesser degree BM but not PB. As invasion by P. vivax was significantly better in reticulocytes generated in vitro, we also suggest that P. vivax may have a preference for invading immature reticulocytes, which should be confirmed in future studies.

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“…Potential that bone marrow could be the source for reticulocytes for in vitro cell culture is becoming an increasingly important area of P. vivax research. Fernandez-Becerra et al have expanded CD34 pos hematopoietic stem cells to generate CD71 pos reticulocytes for in vitro P. vivax culture (63). Others have concentrated their efforts on enriching reticulocytes from cord blood.…”

Section: Discussionmentioning

confidence: 99%

Duffy Antigen Expression in Erythroid Bone Marrow Precursor Cells of Genotypically Duffy Negative Individuals

Dechavanne

Métral

et al. 2018

Preprint

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Tel : +33 (0)1 44 49 31 47 (Dial 0 only within France) SignificanceDuffy blood group negativity results from a single nucleotide polymorphism (SNP) in the gene promoter, and reaches genetic fixation in many African ethnicities. Because the Duffy protein (Fy) is an important contact point during Plasmodium vivax human red blood cell invasion, Fy-negativity is considered to confer resistance to P. vivax malaria.With recent studies in African countries reporting P. vivax infection in Fy-negative people, we studied Fy expression across erythroid development. Here we report that the FY promoter SNP does not abolish Fy protein expression in erythroid progenitors developing in the bone marrow. These results further emphasizes the importance of reticulocytes as targets for P. vivax blood stage infection and propose a mechanism for P. vivax infections in Fy-negative people. Abstract The gene encoding the Duffy blood group protein (Fy, CD234; additional designations Duffy Antigen Receptor of Chemokines [DARC] and Atypical Chemokine Receptor 1 [ACKR1]) is characterized by a SNP in a GATA-1 transcription factor binding site associated with the erythrocyte silent (ES) phenotype. FY ES homozygous people are viewed to be highly resistant to blood stage infection with Plasmodium vivax. Increasingly, however, studies are reporting P.vivax infections in Fy-negative individuals across malarious African countries where FY ES approaches genetic fixation. This suggests that P. vivax has evolved a Fy-independent RBC invasion pathway, or that the GATA-1 SNP does not abolish Fy expression. Here, we tested the second hypothesis through binding studies to erythroid lineage cells using recombinant P. vivax Duffy binding protein, the parasite's invasion ligand and Fy6-specific antibodies. We first observed variable Fy expression on circulating RBCs, irrespective of FY genotype; FY ES RBCs were periodically Fy-positive. Furthermore, during the in vitro erythroid differentiation of CD34+ cells and on ex vivo bone marrow samples, we observed Fy expression on erythroid precursor cells from FY ES people. Finally, the Fy6-specific nanobody, CA111 was used to capture Fy from the surface of FY ES RBCs. Our findings reveal that the GATA-1 SNP does not fully abolish Fy expression and provide insight on potential susceptibility of Fy-negative people to vivax malaria.

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Red blood cells derived from peripheral blood and bone marrow CD34+ human haematopoietic stem cells are permissive to Plasmodium parasites infection

Cited by 15 publications

References 11 publications

CRISPR/Cas9 knockouts reveal genetic interaction between strain-transcendent erythrocyte determinants of Plasmodium falciparum invasion

CRISPR/Cas9 knockouts reveal genetic interaction between strain-transcendent erythrocyte determinants of Plasmodium falciparum invasion

Hematopoietic Stem/Progenitor Cell Sources to Generate Reticulocytes for Plasmodium vivax Culture

Duffy Antigen Expression in Erythroid Bone Marrow Precursor Cells of Genotypically Duffy Negative Individuals

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