Genetic manipulation of cell line derived reticulocytes enables dissection of host malaria invasion requirements

Satchwell, Timothy J.; Wright, Katherine E.; Haydn-Smith, Katy L.; Terán, Fernando Sánchez-Román; Moura, Pedro L.; Hawksworth, Joseph; Frayne, Jan; Toye, Ashley M.; Baum, Jake

doi:10.1038/s41467-019-11790-w

Cited by 23 publications

(33 citation statements)

References 30 publications

(35 reference statements)

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“…As for P. vivax, we are still missing a reliable method for its in vitro culture; the major impediment has been our inability to efficiently handle it under in vitro culture conditions its sole target cell for asexual blood-stage replication: the reticulocyte ( Thomson-Luque et al, 2019). Improved methods for reticulocyte enrichment from different sources have been provided (Vettore et al, 1980;Brun et al, 1990;Kumar et al, 2015;Shaw-Saliba et al, 2016), as well as the production of reticulocytes through better optimized hematopoietic stem cell (HSC) cultures (Giarratana et al, 2005, Noulin et al, 2013Scully et al, 2019) and immortalized lines (Satchwell et al, 2019;Heshusius et al, 2019;Trakarnsanga et al, 2020). The lack of a more efficient enucleation (Menon and Ghaffari, 2021) can be overcome by genetic complementation (Scully et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Home Sweet Home: Plasmodium vivax-Infected Reticulocytes—The Younger the Better?

Thomson-Luque

Bautista

2021

Front. Cell. Infect. Microbiol.

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After a century of constant failure to produce an in vitro culture of the most widespread human malaria parasite Plasmodium vivax, recent advances have highlighted the difficulties to provide this parasite with a healthy host cell to invade, develop, and multiply under in vitro conditions. The actual level of understanding of the heterogeneous populations of cells—framed under the name ‘reticulocytes’—and, importantly, their adequate in vitro progression from very immature reticulocytes to normocytes (mature erythrocytes) is far from complete. The volatility of its individual stability may suggest the reticulocyte as a delusory cell, particularly to be used for stable culture purposes. Yet, the recent relevance gained by a specific subset of highly immature reticulocytes has brought some hope. Very immature reticulocytes are characterized by a peculiar membrane harboring a plethora of molecules potentially involved in P. vivax invasion and by an intracellular complexity dynamically changing upon its quick maturation into normocytes. We analyze the potentialities offered by this youngest reticulocyte subsets as an ideal in vitro host cell for P. vivax.

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Section: Introductionmentioning

confidence: 99%

Home Sweet Home: Plasmodium vivax-Infected Reticulocytes—The Younger the Better?

Thomson-Luque

Bautista

2021

Front. Cell. Infect. Microbiol.

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“…In addition, despite the progress made in generating immortalized erythroid cell-lines retaining a mature phenotype, upscaling the production of these cells for universal usage is challenging 20 – 25 . It is therefore of utmost importance to investigate the contribution of variation in donor erythrocytes in characterizing P. falciparum phenotypic diversity.…”

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confidence: 99%

Blood donor variability is a modulatory factor for P. falciparum invasion phenotyping assays

Thiam

Nyarko

Kusi

et al. 2021

Sci Rep

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Human erythrocytes are indispensable for Plasmodium falciparum development. Unlike other eukaryotic cells, there is no existing erythroid cell line capable of supporting long-term P. falciparum in vitro experiments. Consequently, invasion phenotyping experiments rely on erythrocytes of different individuals. However, the contribution of the erythrocytes variation in influencing invasion rates remains unknown, which represents a challenge for conducting large-scale comparative studies. Here, we used erythrocytes of different blood groups harboring different hemoglobin genotypes to assess the relative contribution of blood donor variability in P. falciparum invasion phenotyping assays. For each donor, we investigated the relationship between parasite invasion phenotypes and erythrocyte phenotypic characteristics, including the expression levels of surface receptors (e.g. the human glycophorins A and C, the complement receptor 1 and decay accelerating factor), blood groups (e.g. ABO/Rh system), and hemoglobin genotypes (e.g. AA, AS and AC). Across all donors, there were significant differences in invasion efficiency following treatment with either neuraminidase, trypsin or chymotrypsin relative to the control erythrocytes. Primarily, we showed that the levels of key erythrocyte surface receptors and their sensitivity to enzyme treatment significantly differed across donors. However, invasion efficiency did not correlate with susceptibility to enzyme treatment or with the levels of the selected erythrocyte surface receptors. Furthermore, we found no relationship between P. falciparum invasion phenotype and blood group or hemoglobin genotype. Altogether, our findings demonstrate the need to consider erythrocyte donor uniformity and anticipate challenges associated with blood donor variability in early stages of large-scale study design.

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“…Furthermore, P. vivax restricted to reticulocytes do not form knobs 68 and completely lack homologs of var genes, offers additional premises to undertake these observations for future research. Evolution of new biological tools such as the ability to engineer and cultivate homogeneous populations of young red blood cells for functional dissection of host factors 69-70 critical for parasite invasion, development and adaptations may contribute to such efforts.…”

Section: Discussionmentioning

confidence: 99%

Reticulocyte Infection Leads to Altered Behaviour, Drug Sensitivity and Host Cell Remodelling byPlasmodium falciparum

Chandramohanadas

Naidu²,

Chu³

et al. 2019

Preprint

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29Plasmodia are host-specific, both at the organism and cellular levels. During asexual 30 development, Plasmodium spp. infect cells of erythroid lineage, with an overall 31 propensity towards reticulocytes. This applies to even Plasmodium (P.) falciparum, the 32 most common causative agent of human malaria, implications of which remain 33 unexplored. Herein, for the first time, we characterize the developmental stages and 34 features of P. falciparum cultured in vitro in young reticulocytes (CD71 + ) in comparison to 35 standard normocyte (CD71 -) cultures. We demonstrate that there are notable differences 36 in the patterns of invasion, development and sensitivity to potent antimalarials (such as 37 artemisinin and dihydroartemisinin) for parasites residing in CD71 + reticulocytes. 38Through a transcriptomic approach, we report that P. falciparum parasites are able to 39 sense the host cell environment, and calibrate their metabolic and host cell remodelling 40 pathways through differential gene expression. These results form an exciting avenue on 41 which hitherto unexplored interactions between Plasmodium spp and different stages of 42 host red blood cells could be investigated in the broader contexts of drug resistance, 43 host tropism and zoonosis. 45Author Summary 46Parasites causing malaria infect red blood cells for development and proliferation during 47 asexual development. This asexual erythrocytic stage determines higher parasite 48 densities and eventual disease manifestation. Although the most virulent species of 49 Plasmodium infecting humans known as Plasmodium falciparum is able to infect red 50 blood cells of all ages, these parasites show a preference for younger blood cells. Of 51 note, the biochemical and biophysical properties of young and adult red blood cells vary 52 significantly. Herein, we undertook a comparative profiling of invasion process, parasite 53 development and drug response of Plasmoddium falciparum in two host cells: young red 54 blood cells (reticulocytes) and mature red blood cells (normocytes). We demonstrate that 55 P. falciparum infects human reticulocytes with higher affinity and demonstrate differential 56 3 sensitivity to drugs such as artemisinin while they reside within reticulocytes. 57Furthermore, we show that P. falciparum is able to detect differences in host 58 environment and adapt to it by changing the expression of genes required for host cell 59 remodelling. 61 Introduction 62Plasmodium infection and associated mortality remain an important concern to the 63 developing world with 218 million malaria cases and ~450,000 deaths annually 1 64 (https://www.who.int/malaria/publications/world-malaria-report-2018/en/). 65Widespread drug-resistance 2-3 and evolution of newer phenotypes; influenced by factors 66 such as changing availability and distribution of insect vector 4 , haematological 67 malignancies(1, 2) 5-6 (thalassemia, sickle cell anaemia, G6PD deficiency etc), providing 68 protective immunity to certain populations 7 , adversely impact malaria eradic...

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Genetic manipulation of cell line derived reticulocytes enables dissection of host malaria invasion requirements

Cited by 23 publications

References 30 publications

Home Sweet Home: Plasmodium vivax-Infected Reticulocytes—The Younger the Better?

Home Sweet Home: Plasmodium vivax-Infected Reticulocytes—The Younger the Better?

Blood donor variability is a modulatory factor for P. falciparum invasion phenotyping assays

Reticulocyte Infection Leads to Altered Behaviour, Drug Sensitivity and Host Cell Remodelling byPlasmodium falciparum

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