Abstract
A feature of the virulent malaria parasite, Plasmodium falciparum, is the sequestration of infected erythrocytes (IEs) to host endothelium. The IEs sequester in the microvasculature by adhesion to host cells resulting in the obstruction of blood flow and often harmful consequences in the host. IEs bind to receptors on host cells with the P. falciparum erythrocyte membrane protein 1 (PfEMP1) that is expressed on the surface of the IEs. The study of parasite cytoadhesion is essential to decipher these ligands, including types of PfEMP1 required for cytoadhesion, the receptors the IEs bind, and how they may be related to the type of malaria disease. An assay for IE adhesion to host cells, including the inhibition of cytoadhesion is described here. The assay involves the purification of IEs with knobs and binding of the IEs to a monolayer of host cells under static conditions. Compounds including proteins, antibodies or drugs can be tested for cytoadhesion inhibitory activity in the assay.
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References
World Health Organization (2020) World malaria report 2020: 20 years of global progress and challenges. World Health Organization, Geneva
Birbeck GL, Molyneux ME, Kaplan PW et al (2010) Blantyre malaria project epilepsy study (BMPES) of neurological outcomes in retinopathy-positive paediatric cerebral malaria survivors: a prospective cohort study. Lancet Neurol 9:1173–1181. https://doi.org/10.1016/S1474-4422(10)70270-2
Milner DA, Whitten RO, Kamiza S et al (2014) The systemic pathology of cerebral malaria in African children. Front Cell Infect Microbiol 4:104
Lee WC, Russell B, Rénia L (2019) Sticking for a cause: the falciparum malaria parasites cytoadherence paradigm. Front Immunol 10:1–15. https://doi.org/10.3389/fimmu.2019.01444
Dondorp AM, Ince C, Charunwatthana P et al (2008) Direct in vivo assessment of microcirculatory dysfunction in severe falciparum malaria. J Infect Dis 197:79–84
Idro R, Kakooza-Mwesige A, Balyejjussa S et al (2010) Severe neurological sequelae and behaviour problems after cerebral malaria in Ugandan children. BMC Res Notes 3:104
Baruch DI, Pasloske BL, Singh HB et al (1995) Cloning the P. falciparum gene encoding PfEMP1, a malarial variant antigen and adherence receptor on the surface of parasitized human erythrocytes. Cell 82:77–87
Hviid L, Jensen ATR (2015) PfEMP1 - a parasite protein family of key importance in plasmodium falciparum malaria immunity and pathogenesis. Adv Parasitol 88:51–84
Udeinya IJ, Schmidt JA, Aikawa M et al (1981) Falciparum malaria infected erythrocytes specifically bind to cultured human endothelial cells. Science 213:555–557
Langreth SG, Peterson E (1985) Pathogenicity, stability, and immunogenicity of a knobless clone of plasmodium falciparum in Colombian owl monkeys. Infect Immun 47:760–766
Claessens A, Adams Y, Ghumra A et al (2012) A subset of group A-like var genes encodes the malaria parasite ligands for binding to human brain endothelial cells. Proc Natl Acad Sci U S A 109:E1772–E1781. https://doi.org/10.1073/pnas.1120461109
Waterkeyn JG, Wickham ME, Davern KM et al (2000) Targeted mutagenesis of plasmodium falciparum erythrocyte membrane protein 3 (PfEMP3) disrupts cytoadherence of malaria-infected red blood cells. EMBO J 19:2813–2823. https://doi.org/10.1093/emboj/19.12.2813
Ganguly AK, Ranjan P, Kumar A, Bhavesh NS (2015) Dynamic association of PfEMP1 and KAHRP in knobs mediates cytoadherence during Plasmodium invasion. Sci Rep 5:8617
Shulman CE, Marshall T, Dorman EK et al (2001) Malaria in pregnancy: adverse effects on haemoglobin levels and birthweight in primigravidae and multigravidae. Trop Med Int Health 6:770–778. https://doi.org/10.1046/j.1365-3156.2001.00786.x
Craig AG, Grau GE, Janse C et al (2012) The role of animal models for research on severe malaria. PLoS Pathog 8:e1002401
Duffier Y, Lorthiois A, Cisteró P et al (2016) A humanized mouse model for sequestration of Plasmodium falciparum sexual stages and in vivo evaluation of gametocytidal drugs. Sci Rep 6:1–9. https://doi.org/10.1038/srep35025
Minkah NK, Schafer C, Kappe SHI (2018) Humanized mouse models for the study of human malaria parasite biology, pathogenesis, and immunity. Front Immunol 9:807. https://doi.org/10.3389/fimmu.2018.00807
Sherman IW, Valdez E (1989) In vitro cytoadherence of Plasmodium falciparum-infected erythrocytes to melanoma cells: factors affecting adhesion. Parasitology 98:359–369
Gillrie MR, Avril M, Brazier AJ et al (2015) Diverse functional outcomes of Plasmodium falciparum ligation of EPCR: potential implications for malarial pathogenesis. Cell Microbiol 17:1883–1899
Avril M, Bernabeu M, Benjamin M et al (2016) Interaction between endothelial protein C receptor and intercellular adhesion molecule 1 to mediate binding of Plasmodium falciparum-infected erythrocytes to endothelial cells. MBio 7:e00615–e00616
Azasi Y, Lindergard G, Ghumra A et al (2018) Infected erythrocytes expressing DC13 PfEMP1 differ from recombinant proteins in EPCR-binding function. Proc Natl Acad Sci U S A 115:1063–1068. https://doi.org/10.1073/pnas.1712879115
Barnwell JW, Asch AS, Nachman RL et al (1989) A human 88-kD membrane glycoprotein (CD36) functions in vitro as a receptor for a cytoadherence ligand on Plasmodium falciparum-infected erythrocytes. J Clin Invest 84:765–772
Berendt AR, Simmons DL, Tansey J et al (1989) Intercellular adhesion molecule-1 is an endothelial cell adhesion receptor for Plasmodium falciparum. Nature 341:57–59
Turner L, Lavstsen T, Berger SS et al (2013) Severe malaria is associated with parasite binding to endothelial protein C receptor. Nature 498:502–505
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Azasi, Y. (2022). Assay of Static Adhesion of Plasmodium falciparum-Infected Erythrocytes to Cells, Including Inhibition of the Adhesion. In: Jensen, A.T.R., Hviid, L. (eds) Malaria Immunology. Methods in Molecular Biology, vol 2470. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2189-9_39
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DOI: https://doi.org/10.1007/978-1-0716-2189-9_39
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