Dielectric characterization of
            Plasmodium falciparum
            -infected red blood cells using microfluidic impedance cytometry

Although malaria is the world's most life-threatening parasitic disease, there is no clear understanding of how certain biophysical properties of infected cells change during the malaria infection cycle. In this article, we use microfluidic impedance cytometry to measure the dielectric properties of Plasmodium falciparum -infected red blood cells ( i- RBCs) at specific time points during the infection cycle. Individual parasites were identified within i- RBCs using green fluorescent protein (GFP) emission. The dielectric properties of cell sub-populations were determined using the multi-shell model. Analysis showed that the membrane capacitance and cytoplasmic conductivity of i- RBCs increased along the infection time course, due to membrane alterations caused by parasite infection. The volume ratio occupied by the parasite was estimated to vary from less than 10% at earlier stages, to approximately 90% at later stages. This knowledge could be used to develop new label-free cell sorting techniques for sample pre-enrichment, improving diagnosis.

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Ultrastructural observations on the in vitro cytoadherence of Plasmodium falciparum infected red blood cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162132 ◽

1990 ◽

Vol 48 (3) ◽

pp. 914-915

Author(s):

D.J.P. Ferguson ◽

A.R. Berendt ◽

J. Tansey ◽

K. Marsh ◽

C.I. Newbold

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Blood Cells ◽

Umbilical Vein ◽

Cell Types ◽

Amelanotic Melanoma ◽

Cytoplasmic Process ◽

Umbilical Vein Endothelial Cells

In human malaria, the most serious clinical manifestation is cerebral malaria (CM) due to infection with Plasmodium falciparum. The pathology of CM is thought to relate to the fact that red blood cells containing mature forms of the parasite (PRBC) cytoadhere or sequester to post capillary venules of various tissues including the brain. This in vivo phenomenon has been studied in vitro by examining the cytoadherence of PRBCs to various cell types and purified proteins. To date, three Ijiost receptor molecules have been identified; CD36, ICAM-1 and thrombospondin. The specific changes in the PRBC membrane which mediate cytoadherence are less well understood, but they include the sub-membranous deposition of electron-dense material resulting in surface deformations called knobs. Knobs were thought to be essential for cytoadherence, lput recent work has shown that certain knob-negative (K-) lines can cytoadhere. In the present study, we have used electron microscopy to re-examine the interactions between K+ PRBCs and both C32 amelanotic melanoma cells and human umbilical vein endothelial cells (HUVEC).We confirm previous data demonstrating that C32 cells possess numerous microvilli which adhere to the PRBC, mainly via the knobs (Fig. 1). In contrast, the HUVEC were relatively smooth and the PRBCs appeared partially flattened onto the cell surface (Fig. 2). Furthermore, many of the PRBCs exhibited an invagination of the limiting membrane in the attachment zone, often containing a cytoplasmic process from the endothelial cell (Fig. 2).

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Endothelin-1 production by a microvascular endothelial cell line treated with Plasmodium falciparum parasitized red blood cells

Clinical Science ◽

10.1042/cs103s464s ◽

2002 ◽

Vol 103 (s2002) ◽

pp. 464S-466S ◽

Cited By ~ 8

Author(s):

Nicoletta BASILICO ◽

Livianna SPECIALE ◽

Silvia PARAPINI ◽

Pasquale FERRANTE ◽

Donatella TARAMELLI

Keyword(s):

Plasmodium Falciparum ◽

Endothelial Cell ◽

Red Blood Cells ◽

Cell Line ◽

Blood Cells ◽

Microvascular Endothelial Cell ◽

Microvascular Endothelium ◽

Endothelial Cell Line ◽

Endothelin 1 ◽

Microvascular Endothelial

In this study, we investigated the production of endothelin 1 (ET-1) by a human microvascular endothelial cell line, HMEC-1, co-cultured with Plasmodium falciparum-parasitized red blood cells (pRBCs). The results indicate that hypoxia increased the basal level of ET-1 production by HMEC-1 cells after 24 or 48h of treatment. However, the co-incubation of HMEC-1 cells with pRBCs, but not with uninfected RBCs, induced a dose-dependent decrease of both constitutive and hypoxia-induced ET-1 production. The inhibition was not due to a decrease in cell viability, as lactate dehydrogenase release remained constant. These results indicate that pRBCs are able to interfere with both the constitutive and stimulated ET-1 release from the microvascular endothelium, thus inducing local modifications of the vascular tone and of the inflammatory response. This could be of relevance in the pathogenesis of the most severe forms of P. falciparum infections, such as cerebral malaria or malaria during pregnancy.

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Determination of quinine in serum, plasma, red blood cells and whole blood in healthy and Plasmodium falciparum malaria cases by high-performance liquid chromatography

Journal of Chromatography B Biomedical Sciences and Applications ◽

10.1016/0378-4347(93)80226-t ◽

1993 ◽

Vol 614 (1) ◽

pp. 87-93 ◽

Cited By ~ 21

Author(s):

Virendra K. Dua ◽

Reema Sarin ◽

Anil Prakash

Keyword(s):

Plasmodium Falciparum ◽

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Red Blood Cells ◽

Falciparum Malaria ◽

Whole Blood ◽

Blood Cells ◽

High Performance ◽

Plasmodium Falciparum Malaria

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The Sites of Sequestration of the Uganda-Palo Alto Strain of Plasmodium falciparum-Infected Red Blood Cells in the Squirrel Monkey, Saimiri sciureus

Journal of Parasitology ◽

10.2307/3278382 ◽

1974 ◽

Vol 60 (3) ◽

pp. 534 ◽

Cited By ~ 10

Author(s):

Henry N. Fremount ◽

Richard N. Rossan

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Squirrel Monkey ◽

Blood Cells ◽

Saimiri Sciureus ◽

Palo Alto

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Red blood cells release microparticles containing human argonaute 2 and miRNAs to target genes of Plasmodium falciparum

Emerging Microbes & Infections ◽

10.1038/emi.2017.63 ◽

2017 ◽

Vol 6 (1) ◽

pp. 1-11 ◽

Cited By ~ 18

Author(s):

Zhensheng Wang ◽

Juemin Xi ◽

Xiao Hao ◽

Weiwei Deng ◽

Juan Liu ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Blood Cells ◽

Target Genes ◽

Argonaute 2

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A Profound Membrane Reorganization Defines Susceptibility of Plasmodium falciparum Infected Red Blood Cells to Lysis by Granulysin and Perforin

Frontiers in Immunology ◽

10.3389/fimmu.2021.643746 ◽

2021 ◽

Vol 12 ◽

Author(s):

Maria Andrea Hernández-Castañeda ◽

Marilyne Lavergne ◽

Pierina Casanova ◽

Bryan Nydegger ◽

Carla Merten ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Blood Cells ◽

Molecular Mechanisms ◽

Γδ T Cells ◽

Parasite Growth ◽

Complex Life Cycle ◽

Cholesterol Depletion ◽

Host Cell Membrane ◽

Pore Forming Proteins

Malaria remains one of the most serious health problems in developing countries. The causative agent of malaria, Plasmodium spp., have a complex life cycle involving multiple developmental stages as well as different morphological, biochemical and metabolic requirements. We recently found that γδ T cells control parasite growth using pore-forming proteins to deliver their cytotoxic proteases, the granzymes, into blood residing parasites. Here, we follow up on the molecular mechanisms of parasite growth inhibition by human pore-forming proteins. We confirm that Plasmodium falciparum infection efficiently depletes the red blood cells of cholesterol, which renders the parasite surrounding membranes susceptible to lysis by prokaryotic membrane disrupting proteins, such as lymphocytic granulysin or the human cathelicidin LL-37. Interestingly, not the cholesterol depletion but rather the simultaneous exposure of phosphatidylserine, a negatively charged phospholipid, triggers resistance of late stage parasitized red blood cells towards the eukaryotic pore forming protein perforin. Overall, by revealing the molecular events we establish here a pathogen-host interaction that involves host cell membrane remodeling that defines the susceptibility towards cytolytic molecules.

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Merocyanine 540-sensitized photoinactivation of human erythrocytes parasitized by Plasmodium falciparum

Blood ◽

10.1182/blood.v80.1.21.bloodjournal80121 ◽

1992 ◽

Vol 80 (1) ◽

pp. 21-24 ◽

Cited By ~ 1

Author(s):

OM Smith ◽

SA Dolan ◽

JA Dvorak ◽

TE Wellems ◽

F Sieber

Keyword(s):

Stem Cells ◽

Plasmodium Falciparum ◽

Hematopoietic Stem Cells ◽

Red Blood Cells ◽

Human Blood ◽

Blood Cells ◽

Potential Usefulness ◽

Hematopoietic Stem ◽

Infected Blood ◽

Merocyanine 540

The purpose of this study was to evaluate the photosensitizing dye merocyanine 540 (MC540) as a means for extracorporeal purging of Plasmodium falciparum-infected erythrocytes from human blood. Parasitized red blood cells bound more dye than nonparasitized cells, and exposure to MC540 and light under conditions that are relatively well tolerated by normal erythrocytes and normal pluripotent hematopoietic stem cells reduced the concentration of parasitized cells by as much as 1,000-fold. Cells parasitized by the chloroquine- sensitive HB3 clone and the chloroquine-resistant Dd2 clone of P falciparum were equally susceptible to MC540-sensitized photolysis. These data suggest the potential usefulness of MC540 in the purging of P falciparum-infected blood.

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PIEZO1 and the mechanism of the long circulatory longevity of human red blood cells

PLoS Computational Biology ◽

10.1371/journal.pcbi.1008496 ◽

2021 ◽

Vol 17 (3) ◽

pp. e1008496

Author(s):

Simon Rogers ◽

Virgilio L. Lew

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Time Course ◽

Clinical Manifestations ◽

Cell Subpopulation ◽

Unexpected Finding ◽

Human Red Blood Cells ◽

Acceptable Model ◽

New Finding ◽

Extended Longevity

Human red blood cells (RBCs) have a circulatory lifespan of about four months. Under constant oxidative and mechanical stress, but devoid of organelles and deprived of biosynthetic capacity for protein renewal, RBCs undergo substantial homeostatic changes, progressive densification followed by late density reversal among others, changes assumed to have been harnessed by evolution to sustain the rheological competence of the RBCs for as long as possible. The unknown mechanisms by which this is achieved are the subject of this investigation. Each RBC traverses capillaries between 1000 and 2000 times per day, roughly one transit per minute. A dedicated Lifespan model of RBC homeostasis was developed as an extension of the RCM introduced in the previous paper to explore the cumulative patterns predicted for repetitive capillary transits over a standardized lifespan period of 120 days, using experimental data to constrain the range of acceptable model outcomes. Capillary transits were simulated by periods of elevated cell/medium volume ratios and by transient deformation-induced permeability changes attributed to PIEZO1 channel mediation as outlined in the previous paper. The first unexpected finding was that quantal density changes generated during single capillary transits cease accumulating after a few days and cannot account for the observed progressive densification of RBCs on their own, thus ruling out the quantal hypothesis. The second unexpected finding was that the documented patterns of RBC densification and late reversal could only be emulated by the implementation of a strict time-course of decay in the activities of the calcium and Na/K pumps, suggestive of a selective mechanism enabling the extended longevity of RBCs. The densification pattern over most of the circulatory lifespan was determined by calcium pump decay whereas late density reversal was shaped by the pattern of Na/K pump decay. A third finding was that both quantal changes and pump-decay regimes were necessary to account for the documented lifespan pattern, neither sufficient on their own. A fourth new finding revealed that RBCs exposed to levels of PIEZO1-medited calcium permeation above certain thresholds in the circulation could develop a pattern of early or late hyperdense collapse followed by delayed density reversal. When tested over much reduced lifespan periods the results reproduced the known circulatory fate of irreversible sickle cells, the cell subpopulation responsible for vaso-occlusion and for most of the clinical manifestations of sickle cell disease. Analysis of the results provided an insightful new understanding of the mechanisms driving the changes in RBC homeostasis during circulatory aging in health and disease.

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