The protein family TcTASV-C is a novel Trypanosoma cruzi virulence factor secreted in extracellular vesicles by trypomastigotes and highly expressed in bloodstream forms

Caeiro, Lucas; Soto, Catalina Dirney Alba; Rizzi, Mariana; Solana, María Elisa; Rodriguez, Giselle; Chidichimo, Agustina; Rodríguez, Matías Exequiel; Sánchez, Daniel O.; Levy, Gabriela Vanesa; Tekiel, Valeria

doi:10.1371/journal.pntd.0006475

Cited by 24 publications

(43 citation statements)

References 61 publications

(89 reference statements)

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“…The interaction between T. cruzi released EVs and target cells may modulate the host responses against the parasite (Cestari et al, 2012;Ramirez et al, 2017). Similarly, EVs obtained from T. cruzi tissue-culture cell-derived trypomastigotes promote functional changes in host-target cells that enhance its infection (Retana Moreira et al, 2019), as well as to modulate the host immune response in favor of the parasite and carry different virulence factors (Ramirez et al, 2017;Caeiro et al, 2018;Lovo-Martins et al, 2018). In line with these reports, we were able to demonstrate that EVs from free trypomastigotes, but not from intracellular amastigotes, were able to increase the host cellular and mitochondrial ROS level which consequently enhanced T. cruzi infection in astrocytes, in a concentration-dependent manner.…”

Section: Discussionmentioning

confidence: 99%

“…Free trypomastigotes of T. cruzi and human astrocytes were washed with serum-free DMEM. Subsequently, secretion equivalents were obtained from 10 8 parasites and 10 7 astrocytes as was previously described with modifications (Caeiro et al, 2018). For this purpose, such parasite and human cells were incubated in DMEM 10% Gibco Exosome-depleted fetal bovine serum (FBS) at 37°C for 6 h in a humidified atmosphere with 5% CO 2 .…”

Section: Isolation Quantification and Characterization Of Extracellmentioning

confidence: 99%

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Priming Astrocytes With HIV-Induced Reactive Oxygen Species Enhances Their Trypanosoma cruzi Infection

et al. 2020

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Introduction: Trypanosoma cruzi is an intracellular protozoa and etiological agent that causes Chagas disease. Its presence among the immunocompromised HIV-infected individuals is relevant worldwide because of its impact on the central nervous system (CNS) causing severe meningoencephalitis. The HIV infection of astrocytes-the most abundant cells in the brain, where the parasite can also be hosted-being able to modify reactive oxygen species (ROS) could influence the parasite growth. In such interaction, extracellular vesicles (EVs) shed from trypomastigotes may alter the surrounding environment including its pro-oxidant status. Methods: We evaluated the interplay between both pathogens in human astrocytes and its consequences on the host cell pro-oxidant condition self-propitiated by the parasiteusing its EVs-or by HIV infection. For this goal, we challenged cultured human primary astrocytes with both pathogens and the efficiency of infection and multiplication were measured by microscopy and flow cytometry and parasite DNA quantification. Mitochondrial and cellular ROS levels were measured by flow cytometry in the presence or not of scavengers with a concomitant evaluation of the cellular apoptosis level. Results: We observed that increased mitochondrial and cellular ROS production boosted significantly T. cruzi infection and multiplication in astrocytes. Such oxidative condition was promoted by free trypomastigotes-derived EVs as well as by HIV infection. Conclusions: The pathogenesis of the HIV-T. cruzi coinfection in astrocytes leads to an oxidative misbalance as a key mechanism, which exacerbates ROS generation and promotes positive feedback to parasite growth in the CNS.

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Section: Discussionmentioning

confidence: 99%

Section: Isolation Quantification and Characterization Of Extracellmentioning

confidence: 99%

Priming Astrocytes With HIV-Induced Reactive Oxygen Species Enhances Their Trypanosoma cruzi Infection

et al. 2020

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“…T. cruzi derived conditioned medium (CM) was obtained by a previously standardized protocol [21]. In brief, cell-derived trypomastigotes (100×10 6 ) were washed with PBS, resuspended in 1 ml of MEM without serum (or MEM without parasites as control medium) and incubated for 6 h at 37°C in a 5% CO 2 humidified atmosphere.…”

Section: Methodsmentioning

confidence: 99%

Transmigration of Trypanosoma cruzi trypomastigotes through 3D cultures resembling a physiological environment

Rodríguez

Rizzi

Caeiro

et al. 2019

Preprint

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AbstractChaga’ disease, caused by the kinetoplastid parasite Trypanosoma cruzi, presents a variety of chronic clinical manifestations whose determinants are still unknown but probably influenced by the host-parasite interplay established during the first stages of the infection, when bloodstream circulating trypomastigotes disseminate to different organs and tissues. After leaving the blood, trypomastigotes must migrate through tissues to invade cells and establish a chronic infection. How this process occurs remains unexplored. Three-dimensional (3D) cultures are physiologically relevant because mimic the microarchitecture of tissues and provide an environment similar to the encountered in natural infections. In this work, we combined the 3D culture technology with host-pathogen interaction, by studying transmigration of trypomastigotes into 3D spheroids. T. cruzi strains with similar infection dynamics in 2D monolayer cultures but with different in vivo behavior (CL Brener, virulent; SylvioX10 no virulent) presented different infection rates in spheroids (CL Brener ∼40%, SylvioX10 <10%). Confocal microscopy images evidenced that trypomastigotes from CL Brener and other highly virulent strains presented a great ability to transmigrate inside 3D spheroids: as soon as 4 hours post infection parasites were found at 50 µm in depth inside the spheroids. CL Brener trypomastigotes were evenly distributed and systematically observed in the space between cells, suggesting a paracellular route of transmigration to deepen into the spheroids. On the other hand, poor virulent strains presented a weak migratory capacity and remained in the external layers of spheroids (<10µm) with a patch-like distribution pattern. The invasiveness -understood as the ability to transmigrate deep into spheroids- was not a transferable feature between strains, neither by soluble or secreted factors nor by co-cultivation of trypomastigotes from invasive and non-invasive strains. We also studied the transmigration of recent T. cruzi isolates from children that were born congenitally infected, which showed a high migrant phenotype while an isolate form an infected mother (that never transmitted the infection to any of her 3 children) was significantly less migratory. Altogether, our results demonstrate that in a 3D microenvironment each strain presents a characteristic migration pattern and distribution of parasites in the spheroids that can be associated to their in vivo behavior. Certainly, the findings presented here could not have been studied with traditional 2D monolayer cultures.Author SummaryTrypanosoma cruzi is the protozoan parasite that causes Chaga’ disease, also known as American trypanosomiasis. Experimental models of the infection evidence that different strains of the parasite present different virulence in the host, which cannot be always reproduced in 2D monolayer cultures. Three dimensional (3D) cultures can be useful models to study complex host-parasite interactions because they mimic in vitro the microarchitecture of tissues and provide an environment similar to the encountered in natural infections. In particular, spheroids are small 3D aggregates of cells that interact with each other and with the extracellular matrix that they secrete resembling the original microenvironment both functionally and structurally. Spheroids have rarely been employed to explore infectious diseases and host-parasite interactions. In this work we studied how bloodstream trypomastigotes transmigrate through 3D spheroids mimicking the picture encountered by parasites in tissues soon after leaving circulation. We showed that the behavior of T. cruzi trypomastigotes in 3D cultures reflects their in vivo virulence: virulent strains transmigrate deeply into spheroids while non-virulent strains remain in the external layers of spheroids. Besides, this work demonstrates the usefulness of 3D cultures as an accurate in vitro model for the study of host-pathogen interactions that could not be addressed with conventional monolayer cultures.

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“…Secreted molecules in PPEVs may also be deployed by a parasite for immune evasion or to avoid extracellular degradation (Caeiro et al, 2018 ). On this basis, EVs from an intracellular and extracellular protozoan parasites promote growth and induce host immune evasion by manipulating the microenvironment for adaptive responses or inhibition of inflammation (Mantel and Marti, 2014 ).…”

Section: Ppevs Bioactive Molecules: Exports and Functionsmentioning

confidence: 99%

Perils and Promises of Pathogenic Protozoan Extracellular Vesicles

Olajide

Cai

2020

Front. Cell. Infect. Microbiol.

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Extracellular vesicles (EVs) are membranous structures formed during biological processes in living organisms. For protozoan parasites, secretion of EVs can occur directly from the parasite organellar compartments and through parasite-infected or antigen-stimulated host cells in response to in vitro and in vivo physiological stressors. These secreted EVs characteristically reflect the biochemical features of their parasitic origin and activating stimuli. Here, we review the species-specific morphology and integrity of parasitic protozoan EVs in concurrence with the origin, functions, and internalization process by recipient cells. The activating stimuli for the secretion of EVs in pathogenic protozoa are discoursed alongside their biomolecules and specific immune cell responses to protozoan parasite-derived EVs. We also present some insights on the intricate functions of EVs in the context of protozoan parasitism.

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The protein family TcTASV-C is a novel Trypanosoma cruzi virulence factor secreted in extracellular vesicles by trypomastigotes and highly expressed in bloodstream forms

Cited by 24 publications

References 61 publications

Priming Astrocytes With HIV-Induced Reactive Oxygen Species Enhances Their Trypanosoma cruzi Infection

Priming Astrocytes With HIV-Induced Reactive Oxygen Species Enhances Their Trypanosoma cruzi Infection

Transmigration of Trypanosoma cruzi trypomastigotes through 3D cultures resembling a physiological environment

Perils and Promises of Pathogenic Protozoan Extracellular Vesicles

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