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Immunomodulatory effect of extracellular vesicles derived from Entamoeba histolytica trophozoites

ABSTRACT: We have isolated and characterized the extracellular vesicles released by Entamoeba histolytica, when alone or in coculture with human neutrophils, and determined their effect on human neutrophil NETs and ROS production.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Entamoeba Histolytica Hm-1:imss-a

TISSUE(S): Blood

SUBMITTER: Cesar Diaz

LAB HEAD: Julio Cesar Carrero

PROVIDER: PXD036228 | Pride | 2023-03-10

REPOSITORIES: Pride

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			Action	DRS
	20211011_1-_1_.mgf	Mgf
	20211011_1.mzML	Mzml
	20211011_1.mzid.gz	Mzid
	20211011_1.raw	Raw
	20211011_2-_1_.mgf	Mgf

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Publications

Immunomodulatory effect of extracellular vesicles from <i>Entamoeba histolytica</i> trophozoites: Regulation of NETs and respiratory burst during confrontation with human neutrophils.

Díaz-Godínez César C Ríos-Valencia Diana G DG García-Aguirre Samuel S Martínez-Calvillo Santiago S Carrero Julio César JC

Frontiers in cellular and infection microbiology 20221028

Parasites release extracellular vesicles (EVs) which, in some cases, modulate the host's immune response contributing to the establishment of the infection. In this work we have isolated and characterized the EVs released by trophozoites of the human protozoan parasite <i>Entamoeba histolytica</i>, the causal agent of amoebiasis, when alone or in coculture with human neutrophils, and determined their effect on neutrophil NETs and ROS production. Nanoparticle tracking analysis showed that amoebic ...[more]

PMID: 36389143

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Gene expression in phagocytic and non-phagocytic Entamoeba histolytica populations

Project description:The ability of Entamoeba histolytica to phagocytose host cells correlates to observed virulence in vivo. To better understand the mechanism of phagocytosis we used paramagnetic beads coated with host ligand and sorted trophozoites based on phagocytic ability. Gene expression was then measured in both the sorted phagocytic and non-phagocytic populations using a custom Affymetrix chip for E. histolytica. Feed forward regulation of phagocytosis by Entamoeba histolytica. Infection and Immunity. PMID 23045476 M280 Streptavidin Dynabeads (Invitrogen) were labeled with 20ug/mL biotinylated Human C1q (Quidel). Entamoeba histolytica (strain HM1) were washed twice with PBS then resuspended with the labeled beads at a 10:1 ratio of beads to trophozoites. Samples were incubated at 37Â°C for 45 minutes, washed twice with agitation to remove adherent beads, then resuspended in MACS buffer. Samples were loaded into magnetic columns (Miltenyi Biotec) and trophozoites were seperated according to manufacturer's protocols. phagocytic vs. non-phagocytic Entamoeba histolytica populations

2012-10-16 | E-GEOD-41604 | biostudies-arrayexpress

Syntrophic coculture of Syntrophobacter fumaroxidans MPOBT and Geobacter sulfurreducens PCA

Project description:Propionate accumulation is an important bottleneck for anaerobic degradation of organic matter. We hypothesized that propionate conversion by a novel coculture of Syntrophobacter fumaroxidans and Geobacter sulfurreducens can be an alternative strategy for propionate oxidation coupled to Fe(III) reduction. In this study, we successfully cocultured S. fumaroxidans and G. sulfurreducens on propionate and Fe(III). Proteomic analyses of this coculture provided insights into the underlying mechanisms of propionate metabolism pathway and interspecies electron transfer mechanism. Our study can be further useful in understanding syntrophic propionate degradation in bioelectrochemical and anaerobic digestion systems.

2022-02-17 | PXD027104 | Pride

Small RNAs during E. histolytica stress response

Project description:We analyzed ~27nt small RNAs from Entamoeba histolytica trophozoites in basal conditions and after heat shock or oxidative stress E. histolytica trophozoites were treated with 1mM H2O2 for 1hr, or heat shocked at 42°C for 1hr and RNA was isolated and small RNA populations were compared to small RNA populations from untreated trophozoites

2015-07-22 | E-GEOD-67230 | biostudies-arrayexpress

Coactosin phosphorylation controls Entamoeba histolytica 1 cell membrane protusions and cell motility

Project description:Invasion of the colon wall by Entamoeba histolytica during amoebic dysentery entails migration of trophozoites through tissue layers that are rich in extracellular matrix (ECM). Transcriptional silencing of the E. histolytica matrix surface metalloprotease EhMSP-1 produces hyper-adherent, less motile trophozoites that are deficient in forming invadosomes. Reversible protein phosphorylation is often implicated in regulation of cell motility and invadosome formation. To identify such intermediaries of the EhMSP-1 silenced phenotype, here we compared the phosphoproteome of EhMSP-1 silenced and vector control trophozoites using quantitative mass spectrometry based proteomics. Six proteins were found to be differentially phosphorylated in EhMSP-1 silenced and control cells, including EhCoactosin, a member of the ADF/Cofilin family of actin binding proteins, which was hyperphosphorylated at serine 147 (S147). Regulated over-expression of wild type, phosphomimetic (S147D), and non phosphorylatable (S147A) EhCoactosin variants was used to test if S147 phosphorylation functions in control of E. histolytica actin dynamics. Each of the over-expressed proteins co37 localized with F-actin during E. histolytica phagocytosis. Nonetheless, trophozoites overexpressing EhCoactosin S147D formed more and poorly coordinated small membrane protrusions compared to control or S147A expressing cells, while trophozoites overexpressing EhCoactosin S147A were significantly more motile within a model of mammalian ECM. Therefore, although EhCoactosin’s actin-binding ability appeared unaffected by S147 phosphorylation, EhCoactosin phosphorylation helps to regulate amoebic motility. These data help to understand the mechanisms underlying altered adherence and motility in EhMSP-1 silenced trophozoites, and lay the groundwork for identifying kinases and phosphatases critical for control of amoebic invasiveness.

2020-07-15 | PXD018276 | Pride

Transcriptional Metronidazole treatment

Project description:This study aims to search for the effect of 50uM Metronidazole against Entamoeba histolytica and thereby elucidating their mechanism of action

2014-01-14 | E-MTAB-1763 | biostudies-arrayexpress

Role of REG 1 in Entamoeba histolytica colitis

Project description:Differential expression was used to access gene differences after Entamoeba histolytica infection. Entamoeba histolytica is an important diarrheal pathogen worldwide, and induces apoptosis of the intestinal epithelium as part of its disease process. Regenerating (REG) 1 protein is anti-apoptotic. We investigated the involvement of REG 1 in E. histolytica colitis. Colonic biopsy samples were obtained from 8 subjects with acute E. histolytica colitis, and again 60 day later during convalescence. Gene expression in the human colon during acute and convalescent E. histolytica disease was evaluated using microarray and confirmed by polymerase chain reaction (PCR). REG 1 protein expression was evaluated with immunohistochemistry. The mechanism of REG 1 involvement in E. histolytica disease was subsequently investigated with a mouse model. REG 1A and REG 1B were the most upregulated genes in the human intestine in acute versus convalescent E. histolytica disease (p=0.003 and p=0.006 respectively). PCR confirmed the microarray results (p=<0.001 and p=0.001 respectively). Increased REG 1A and REG 1B protein expression was similarly observed by immunohistochemistry. REG 1 -/-mice were found to be significantly more susceptible to E. histolytica infection than wild type mice. Intestinal Biopsies were taken at day 1 and day 60 post infection.

2011-05-24 | E-GEOD-23750 | biostudies-arrayexpress

Transcription profiling of human brain endothelial cells treated with TNF then incubated with platelets, P.falciparum-parasited red blood cells or normal red blood cells

Project description:HBEC-5i (human brain endothelial cells) grown to confluence in 6-well tissue culture plates. Cocultures were either left unstimulated or incubated overnight with recombinant TNF?. HBEC were then washed in PBS and incubated with PLT (platelets), pRBC (P.falciparum-parasited red blood cells) or NRBC (normal red blood cells) according to the experimental conditions. Then HBEC were washed three times with PBS, and were harvested at 0 and 5h coculture. Each experimental condition was realized in triplicate.

2009-01-15 | E-MEXP-1989 | biostudies-arrayexpress

Transcriptional profiling of human liver sinusoidal endothelial cells respond to interaction with Entamoeba histolityca.

Project description:incubation for 15 min and 30 min of human liver sinusoidal cell cultures with virulent or virulence-attenuated Entamoeba histolytica or incubation without parasites added

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Project description:The availability of two genetically very similar clones (A and B) derived from the laboratory isolate Entamoeba histolytica HM-1:IMSS, which differ in their virulence properties, provides a powerful tool for identifying pathogenicity factors of the causative agent of human amoebiasis. We isolated RNA from each clone for transcriptional comparison.

2010-01-06 | E-MEXP-2504 | biostudies-arrayexpress

Transcriptional profiling by array of Entamoeba histolytica Mutant Derived from Virulent Strain

Project description:Amoebiasis is a disease caused by the protozoan parasite, Entamoeba histolytica, with or without clinical symptoms . It has been suggested that pathogenic amoebae use three major virulence factors: Gal/GalNAc lectin, cysteine proteinases and amoebapores. However, these molecules cannot exclusively be responsible for amoebic virulence, because most of them are found in pathogenic E. histolytica as well as in non-pathogenic E. dispar, a not close correlation has been found in amoebic strains with different virulence phenotype. Due to the high genetic variability of E. histolytica isolates and strains, differential gene expression might simply reflect inter-isolate genetic variation rather than specific differences linked to virulence. In the current study, we obtained a non-virulent UG10 mutant derived from E. histolytica HM-1:IMSS. Comparing the transcriptome of both strains, no differences in gene expression of the classical virulence factors were observed. RAB family GTPase and AIG1 family members showed higher transcriptional levels in the virulent strain than in the non-virulent mutant. Our study suggests a number of novel gene products in E. histolytica with a role yet to determine in cell physiology and involvement in the pathogenesis process.

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