Using a new three-dimensional CUBIC tissue-clearing method to examine the brain during experimental cerebral malaria

2021 ◽  
Author(s):  
Julia Matsuo-Dapaah ◽  
Michelle Sue Jann Lee ◽  
Ken J Ishii ◽  
Kazuki Tainaka ◽  
Cevayir Coban
Keyword(s):  
Olfactory Bulb ◽  
Cerebral Malaria ◽  
Fluorescent Microscopy ◽  
Three Dimensions ◽  
Whole Brain ◽  
Cerebral Microvessels ◽  
Experimental Cerebral Malaria ◽  
Tissue Clearing ◽  
The Brain ◽  
Clearing Method

Abstract Cerebral malaria (CM) is a life-threatening complication of the malaria disease caused by Plasmodium falciparum infection and is responsible for the death of half a million people annually. The molecular pathogenesis underlying CM in humans is not completely understood, although sequestration of infected erythrocytes in cerebral microvessels is thought to play a major role. In contrast, experimental cerebral malaria (ECM) models in mice have been thought to be distinct from human CM, and are mainly caused by inflammatory mediators. Here, to understand the spatial distribution and the potential sequestration of parasites in the whole-brain microvessels during a mouse model of ECM, we utilized the new tissue-clearing method CUBIC (Clear, Unobstructed, Brain/Body Imaging Cocktails and Computational analysis) with light sheet fluorescent microscopy (LSFM), and reconstructed images in three-dimensions (3D). We demonstrated significantly greater accumulation of Plasmodium berghei ANKA (PbANKA) parasites in the olfactory bulb of mice, compared with the other parts of the brain, including the cerebral cortex, cerebellum, and brainstem. Furthermore, we show that PbANKA parasites preferentially accumulate in the brainstem when the olfactory bulb is surgically removed. This study therefore not only highlights a successful application of CUBIC tissue-clearing technology to visualize the whole brain and its microvessels during ECM, but it also shows CUBIC’s future potential for visualizing pathological events in the whole ECM brain at the cellular level, an achievement that would greatly advance our understanding of human cerebral malaria.

CCR5 deficiency decreases susceptibility to experimental cerebral malaria

Blood ◽  
2003 ◽  
Vol 101 (11) ◽  
pp. 4253-4259 ◽  
Author(s):  
Elodie Belnoue ◽  
Michèle Kayibanda ◽  
Jean-Christophe Deschemin ◽  
Mireille Viguier ◽  
Matthias Mack ◽  
...  
Keyword(s):  
T Cells ◽  
Cerebral Malaria ◽  
Cd8 T Cells ◽  
Cytokine Production ◽  
Wild Type ◽  
Experimental Cerebral Malaria ◽  
Pathologic Features ◽  
Th1 Cytokine ◽  
The Brain ◽  
Deficient Mice

Abstract Infection of susceptible mouse strains with Plasmodium berghei ANKA (PbA) is a valuable experimental model of cerebral malaria (CM). Two major pathologic features of CM are the intravascular sequestration of infected erythrocytes and leukocytes inside brain microvessels. We have recently shown that only the CD8+ T-cell subset of these brain-sequestered leukocytes is critical for progression to CM. Chemokine receptor–5 (CCR5) is an important regulator of leukocyte trafficking in the brain in response to fungal and viral infection. Therefore, we investigated whether CCR5 plays a role in the pathogenesis of experimental CM. Approximately 70% to 85% of wild-type and CCR5+/- mice infected with PbA developed CM, whereas only about 20% of PbA-infected CCR5-deficient mice exhibited the characteristic neurologic signs of CM. The brains of wild-type mice with CM showed significant increases in CCR5+ leukocytes, particularly CCR5+ CD8+ T cells, as well as increases in T-helper 1 (Th1) cytokine production. The few PbA-infected CCR5-deficient mice that developed CM exhibited a similar increase in CD8+ T cells. Significant leukocyte accumulation in the brain and Th1 cytokine production did not occur in PbA-infected CCR5-deficient mice that did not develop CM. Moreover, experiments using bone marrow (BM)–chimeric mice showed that a reduced but significant proportion of deficient mice grafted with CCR5+ BM develop CM, indicating that CCR5 expression on a radiation-resistant brain cell population is necessary for CM to occur. Taken together, these results suggest that CCR5 is an important factor in the development of experimental CM.

scholarly journals IFN-γ–Producing CD4+ T Cells Promote Experimental Cerebral Malaria by Modulating CD8+ T Cell Accumulation within the Brain

2012 ◽  
Vol 189 (2) ◽  
pp. 968-979 ◽  
Author(s):  
Ana Villegas-Mendez ◽  
Rachel Greig ◽  
Tovah N. Shaw ◽  
J. Brian de Souza ◽  
Emily Gwyer Findlay ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cerebral Malaria ◽  
Cd4 T Cells ◽  
Cd8 T Cell ◽  
Experimental Cerebral Malaria ◽  
Cell Accumulation ◽  
Ifn Γ ◽  
The Brain

scholarly journals Cerebral malaria: why experimental murine models are required to understand the pathogenesis of disease

Parasitology ◽  
2009 ◽  
Vol 137 (5) ◽  
pp. 755-772 ◽  
Author(s):  
J. BRIAN de SOUZA ◽  
JULIUS C. R. HAFALLA ◽  
ELEANOR M. RILEY ◽  
KEVIN N. COUPER
Keyword(s):  
Cerebral Malaria ◽  
Malaria Infection ◽  
Clinical Symptoms ◽  
Experimental Models ◽  
Murine Models ◽  
Experimental Cerebral Malaria ◽  
Sequence Of Events ◽  
Life Threatening ◽  
Life Threatening Complication ◽  
The Brain

SUMMARYCerebral malaria is a life-threatening complication of malaria infection. The pathogenesis of cerebral malaria is poorly defined and progress in understanding the condition is severely hampered by the inability to study in detail,ante-mortem, the parasitological and immunological events within the brain that lead to the onset of clinical symptoms. Experimental murine models have been used to investigate the sequence of events that lead to cerebral malaria, but there is significant debate on the merits of these models and whether their study is relevant to human disease. Here we review the current understanding of the parasitological and immunological events leading to human and experimental cerebral malaria, and explain why we believe that studies with experimental models of CM are crucial to define the pathogenesis of the condition.

scholarly journals Requirement for Tumor Necrosis Factor Receptor 2 Expression on Vascular Cells To Induce Experimental Cerebral Malaria

2002 ◽  
Vol 70 (10) ◽  
pp. 5857-5859 ◽  
Author(s):  
Benjamin Stoelcker ◽  
Thomas Hehlgans ◽  
Karin Weigl ◽  
Horst Bluethmann ◽  
Georges E. Grau ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Cerebral Malaria ◽  
Tumor Necrosis ◽  
Tumor Necrosis Factor Receptor ◽  
Experimental Cerebral Malaria ◽  
Brain Vasculature ◽  
Bone Marrow Chimeras ◽  
The Brain ◽  
Necrosis Factor

ABSTRACT Using tumor necrosis factor receptor type 2 (TNFR2)-deficient mice and generating bone marrow chimeras which express TNFR2 on either hematopoietic or nonhematopoietic cells, we demonstrated the requirement for TNFR2 expression on tissue cells to induce lethal cerebral malaria. Thus, TNFR2 on the brain vasculature mediates tumor necrosis factor-induced neurovascular lesions in experimental cerebral malaria.

scholarly journals The mTOR Pathway in Platelets Contributes to the Pathophysiology of Experimental Cerebral Malaria

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 580-580
Author(s):  
Irina Portier ◽  
Frederik Denorme ◽  
Kimberly A Queisser ◽  
Yasuhiro Kosaka ◽  
Aaron C Petrey ◽  
...  
Keyword(s):  
Cerebral Malaria ◽  
Protein Translation ◽  
Mtor Pathway ◽  
Rna Seq ◽  
Wild Type ◽  
Advisory Committees ◽  
Experimental Cerebral Malaria ◽  
Inflammatory Monocytes ◽  
The Brain

Abstract Background: Cerebral malaria is a highly prevalent infectious disease in Sub-Saharan Africa caused by the Plasmodium parasite. The pathogenesis of cerebral malaria results from damaged vascular endothelium induced by parasite sequestration, inflammatory cytokine production and vascular leakage, which results in increased brain permeability and death. While maladaptive responses from immune cells are thought to contribute, growing evidence suggests a crucial role of platelets in malaria pathophysiology. The mammalian target of rapamycin (mTOR) pathway is critical in regulating outcomes in malaria. Previous studies have demonstrated an mTOR specific inhibitor, rapamycin, is protective in a mouse model of experimental cerebral malaria (ECM). However, if the mTOR pathway in platelets specifically contributes to the pathogenesis of malaria is unknown. Methods: Platelet-specific mTOR-deficient (mTOR plt-/-) mice and littermate controls were subjected to a well-established model of ECM, using Plasmodium berghei ANKA. In addition, platelets isolated from human malaria patients were examined for differential regulation of the mTOR pathway using RNA-seq. Results: Platelet RNA-seq and Ingenuity Pathway Analysis from patients infected with P. vivax demonstrated enrichment of mTOR-associated pathways in platelets, such as mTOR signaling and p70S6K signaling, indicating mTOR associated genes are upregulated in human platelets during malaria infection. In mice infected with P. berghei ANKA, the mTOR pathway was activated in bone marrow-megakaryocytes and platelets based on phosphorylation of mTOR and its downstream effector, 4E-BP1. As the mTOR pathway regulates protein translation in platelets, we examined de novo protein synthesis and observed increased protein translation in platelets isolated from mice infected with P. berghei ANKA compared to uninfected controls. To study the specific role of platelet mTOR during ECM pathogenesis, mTOR plt-/- mice and wild-type controls (mTOR plt+/+), were infected with P. berghei ANKA. Platelet deficient-mTOR mice had significantly (p=0.0336) prolonged survival compared to wild-type mice. Increased survival was independent of parasitemia, suggesting platelets did not alter parasite reproduction. While thrombocytopenia and anemia were similar in both genotypes, mTOR plt-/- mice had significantly reduced brain (p=0.0067) and lung (p<0.0001) vascular permeability during late-stage ECM. Interestingly, flow cytometric assessment of leukocyte recruitment to the brain demonstrated a 1.7-fold (p=0.0442) reduction in inflammatory monocytes in platelet-deficient mTOR mice. However, mTOR plt-/- mice had significantly (1.4-fold, p=0.007) more inflammatory monocytes in the blood. Interestingly, circulating platelet-monocytes aggregates were significantly less in mTOR plt-/- compared to mTOR plt+/+ (p=0.0433). Taken together, these results suggest that platelets assist in the recruitment of leukocytes to the brain vasculature during ECM, which is impaired when mTOR is ablated. Conclusions: Our data demonstrates that the mTOR pathway in platelets plays a significant role in malaria pathogenesis. Deletion of platelet mTOR reduces vascular permeability and prolongs survival during ECM. We hypothesize that altered platelet-inflammatory monocyte interactions drive this phenotype. Disclosures Rondina: Platelet Transcriptomics: Patents & Royalties; Acticor Biotech: Membership on an entity's Board of Directors or advisory committees; Platelet Biogenesis: Membership on an entity's Board of Directors or advisory committees; Novartis: Research Funding.

scholarly journals Beta Interferon Suppresses the Development of Experimental Cerebral Malaria

2011 ◽  
Vol 79 (4) ◽  
pp. 1750-1758 ◽  
Author(s):  
Craig N. Morrell ◽  
Kalyan Srivastava ◽  
AnneMarie Swaim ◽  
M. Teresa Lee ◽  
Jun Chen ◽  
...  
Keyword(s):  
T Cell ◽  
Cerebral Malaria ◽  
Blood Brain Barrier ◽  
Vascular Inflammation ◽  
Brain Barrier ◽  
Experimental Cerebral Malaria ◽  
Beta Interferon ◽  
Cxcr3 Expression ◽  
Blood Brain ◽  
The Brain

ABSTRACTCerebral malaria (CM) is a major complication ofPlasmodium falciparuminfection, particularly in children. The pathogenesis of cerebral malaria involves parasitized red blood cell (RBC)-mediated vascular inflammation, immune stimulation, loss of blood-brain barrier integrity, and obstruction of cerebral capillaries. Therefore, blunting vascular inflammation and immune cell recruitment is crucial in limiting the disease course. Beta interferon (IFN-β) has been used in the treatment of diseases, such as multiple sclerosis (MS) but has not yet been explored in the treatment of CM. Therefore, we sought to determine whether IFN-β also limits disease progression in experimental cerebral malaria (ECM).Plasmodium berghei-infected mice treated with IFN-β died later and showed increased survival, with improved blood-brain barrier function, compared to infected mice. IFN-β did not alter systemic parasitemia. However, we identified multiple action sites that were modified by IFN-β administration.P. bergheiinfection resulted in increased expression of chemokine (C-X-C motif) ligand 9 (CXCL9) in brain vascular endothelial cells that attract T cells to the brain, as well as increased T-cell chemokine (C-X-C motif) receptor 3 (CXCR3) expression. The infection also increased the cellular content of intercellular adhesion molecule 1 (ICAM-1), a molecule important for attachment of parasitized RBCs to the endothelial cell. In this article, we report that IFN-β treatment leads to reduction of CXCL9 and ICAM-1 in the brain, reduction of T-cell CXCR3 expression, and downregulation of serum tumor necrosis factor alpha (TNF-α). In addition, IFN-β-treatedP. berghei-infected mice also had fewer brain T-cell infiltrates, further demonstrating its protective effects. Hence, IFN-β has important anti-inflammatory properties that ameliorate the severity of ECM and prolong mouse survival.

scholarly journals Proteomic profiling of the brain of mice with experimental cerebral malaria

2018 ◽  
Vol 180 ◽  
pp. 61-69 ◽  
Author(s):  
Ehab Moussa ◽  
Honglei Huang ◽  
Malika Ahras ◽  
Amar Lall ◽  
Marie L. Thezenas ◽  
...  
Keyword(s):  
Cerebral Malaria ◽  
Proteomic Profiling ◽  
Experimental Cerebral Malaria ◽  
The Brain

scholarly journals Assessing Vascular Permeability during Experimental Cerebral Malaria by a Radiolabeled Monoclonal Antibody Technique

2001 ◽  
Vol 69 (5) ◽  
pp. 3460-3465 ◽  
Author(s):  
Henri C. van der Heyde ◽  
Philippe Bauer ◽  
Guang Sun ◽  
Wun-Ling Chang ◽  
Lijia Yin ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cerebral Malaria ◽  
Vascular Permeability ◽  
Plasmodium Berghei ◽  
Blue Dye ◽  
Vascular Endothelial ◽  
Antibody Technique ◽  
Experimental Cerebral Malaria ◽  
Endothelial Integrity ◽  
The Brain

ABSTRACT Vascular endothelial integrity, assessed by Evans blue dye extrusion and radiolabeled monoclonal antibody leakage, was markedly compromised in the brain, lung, kidney, and heart duringPlasmodium berghei infection, a well-recognized model for human cerebral malaria. The results for vascular permeability from both methods were significantly (P < 0.001) related.

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