Thrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence

ABSTRACT Plasmodium falciparum malaria remains one of the most deadly infections worldwide. The pathogenesis of the infection results from the sequestration of infected erythrocytes (IRBC) in vital organs, including the brain, with resulting impairment of blood flow, hypoxia, and lactic acidosis. Se...

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Autores principales:	Mark R. Gillrie, Bernard Renaux, Eleanor Russell-Goldman, Marion Avril, Andrew J. Brazier, Koichiro Mihara, Enrico Di Cera, Danny A. Milner, Morley D. Hollenberg, Joseph D. Smith, May Ho
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Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ef9c28ad329b4fbba481ab5d041f6224
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spelling	oai:doaj.org-article:ef9c28ad329b4fbba481ab5d041f62242021-11-15T15:50:14ZThrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence10.1128/mBio.01120-162150-7511https://doaj.org/article/ef9c28ad329b4fbba481ab5d041f62242016-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01120-16https://doaj.org/toc/2150-7511ABSTRACT Plasmodium falciparum malaria remains one of the most deadly infections worldwide. The pathogenesis of the infection results from the sequestration of infected erythrocytes (IRBC) in vital organs, including the brain, with resulting impairment of blood flow, hypoxia, and lactic acidosis. Sequestration occurs through the adhesion of IRBC to host receptors on microvascular endothelium by Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1), a large family of variant surface antigens, each with up to seven extracellular domains that can bind to multiple host receptors. Consequently, antiadhesive therapies directed at single endothelial adhesion molecules may not be effective. In this study, we demonstrated that the serine protease thrombin, which is pivotal in the activation of the coagulation cascade, cleaved the major parasite adhesin on the surface of IRBC. As a result, adhesion under flow was dramatically reduced, and already adherent IRBC were detached. Thrombin cleavage sites were mapped to the Duffy binding-like δ1 (DBLδ1) domain and interdomains 1 and 2 in the PfEMP1 of the parasite line IT4var19. Furthermore, we observed an inverse correlation between the presence of thrombin and IRBC in cerebral malaria autopsies of children. We investigated a modified (R67A) thrombin and thrombin inhibitor, hirugen, both of which inhibit the binding of substrates to exosite I, thereby reducing its proinflammatory properties. Both approaches reduced the barrier dysfunction induced by thrombin without affecting its proteolytic activity on PfEMP1, raising the possibility that thrombin cleavage of variant PfEMP1 may be exploited as a broadly inhibitory antiadhesive therapy. IMPORTANCE Plasmodium falciparum malaria is the third leading cause of mortality due to a pathogen, with 214 million people infected and 438,000 deaths annually. The adhesion of Plasmodium falciparum-infected erythrocytes (IRBC) to microvascular endothelium is a major pathological process in severe malaria. While the recent implementation of artemisinin-based antimalarial therapy for severe malaria improves patient survival by targeting all parasite stages, antiparasite drugs alone may not immediately reverse pathophysiological processes in occluded vessels. Here we show that thrombin, an enzyme intimately involved in the clotting process, cleaves the main parasite adhesin expressed on the surface of IRBC, thereby preventing and reversing the binding of IRBC to endothelial cells. This beneficial effect of thrombin can be achieved by modified thrombins that cause significantly less clotting and vessel leakage while preserving the ability to cleave the parasite protein. Our results provide the basis for using modified thrombins as adjunctive therapy in severe malaria.Mark R. GillrieBernard RenauxEleanor Russell-GoldmanMarion AvrilAndrew J. BrazierKoichiro MiharaEnrico Di CeraDanny A. MilnerMorley D. HollenbergJoseph D. SmithMay HoAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 7, Iss 5 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Mark R. Gillrie Bernard Renaux Eleanor Russell-Goldman Marion Avril Andrew J. Brazier Koichiro Mihara Enrico Di Cera Danny A. Milner Morley D. Hollenberg Joseph D. Smith May Ho Thrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence
description	ABSTRACT Plasmodium falciparum malaria remains one of the most deadly infections worldwide. The pathogenesis of the infection results from the sequestration of infected erythrocytes (IRBC) in vital organs, including the brain, with resulting impairment of blood flow, hypoxia, and lactic acidosis. Sequestration occurs through the adhesion of IRBC to host receptors on microvascular endothelium by Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1), a large family of variant surface antigens, each with up to seven extracellular domains that can bind to multiple host receptors. Consequently, antiadhesive therapies directed at single endothelial adhesion molecules may not be effective. In this study, we demonstrated that the serine protease thrombin, which is pivotal in the activation of the coagulation cascade, cleaved the major parasite adhesin on the surface of IRBC. As a result, adhesion under flow was dramatically reduced, and already adherent IRBC were detached. Thrombin cleavage sites were mapped to the Duffy binding-like δ1 (DBLδ1) domain and interdomains 1 and 2 in the PfEMP1 of the parasite line IT4var19. Furthermore, we observed an inverse correlation between the presence of thrombin and IRBC in cerebral malaria autopsies of children. We investigated a modified (R67A) thrombin and thrombin inhibitor, hirugen, both of which inhibit the binding of substrates to exosite I, thereby reducing its proinflammatory properties. Both approaches reduced the barrier dysfunction induced by thrombin without affecting its proteolytic activity on PfEMP1, raising the possibility that thrombin cleavage of variant PfEMP1 may be exploited as a broadly inhibitory antiadhesive therapy. IMPORTANCE Plasmodium falciparum malaria is the third leading cause of mortality due to a pathogen, with 214 million people infected and 438,000 deaths annually. The adhesion of Plasmodium falciparum-infected erythrocytes (IRBC) to microvascular endothelium is a major pathological process in severe malaria. While the recent implementation of artemisinin-based antimalarial therapy for severe malaria improves patient survival by targeting all parasite stages, antiparasite drugs alone may not immediately reverse pathophysiological processes in occluded vessels. Here we show that thrombin, an enzyme intimately involved in the clotting process, cleaves the main parasite adhesin expressed on the surface of IRBC, thereby preventing and reversing the binding of IRBC to endothelial cells. This beneficial effect of thrombin can be achieved by modified thrombins that cause significantly less clotting and vessel leakage while preserving the ability to cleave the parasite protein. Our results provide the basis for using modified thrombins as adjunctive therapy in severe malaria.
format	article
author	Mark R. Gillrie Bernard Renaux Eleanor Russell-Goldman Marion Avril Andrew J. Brazier Koichiro Mihara Enrico Di Cera Danny A. Milner Morley D. Hollenberg Joseph D. Smith May Ho
author_facet	Mark R. Gillrie Bernard Renaux Eleanor Russell-Goldman Marion Avril Andrew J. Brazier Koichiro Mihara Enrico Di Cera Danny A. Milner Morley D. Hollenberg Joseph D. Smith May Ho
author_sort	Mark R. Gillrie
title	Thrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence
title_short	Thrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence
title_full	Thrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence
title_fullStr	Thrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence
title_full_unstemmed	Thrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence
title_sort	thrombin cleavage of <named-content content-type="genus-species">plasmodium falciparum</named-content> erythrocyte membrane protein 1 inhibits cytoadherence
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/ef9c28ad329b4fbba481ab5d041f6224
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Thrombin Cleavage of <named-content content-type="genus-species">Plasmodium falciparum</named-content> Erythrocyte Membrane Protein 1 Inhibits Cytoadherence

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