Proteomic analysis reveals different composition of extracellular vesicles released by two Trypanosoma cruzi strains associated with their distinct interaction with host cells

Abstract

Trypanosoma cruzi, the aetiologic agent of Chagas disease, releases vesicles containing a wide range of surface molecules known to affect the host immunological responses and the cellular infectivity. Here, we compared the secretome of two distinct strains (Y and YuYu) of T. cruzi, which were previously shown to differentially modulate host innate and acquired immune responses. Tissue culture-derived trypomastigotes of both strains secreted extracellular vesicles (EVs), as demonstrated by electron scanning microscopy. EVs were purified by exclusion chromatography or ultracentrifugation and quantitated using nanoparticle tracking analysis. Trypomastigotes from YuYu strain released higher number of EVs than those from Y strain, enriched with virulence factors trans-sialidase (TS) and cruzipain. Proteomic analysis confirmed the increased abundance of proteins coded by the TS gene family, mucin-like glycoproteins, and some typical exosomal proteins in the YuYu strain, which also showed considerable differences between purified EVs and vesicle-free fraction as compared to the Y strain. To evaluate whether such differences were related to parasite infectivity, J774 macrophages and LLC-MK2 kidney cells were preincubated with purified EVs from both strains and then infected with Y strain trypomastigotes. EVs released by YuYu strain caused a lower infection but higher intracellular proliferation in J774 macrophages than EVs from Y strain. In contrast, YuYu strain-derived EVs caused higher infection of LLC-MK2 cells than Y strain-derived EVs. In conclusion, quantitative and qualitative differences in EVs and secreted proteins from different T. cruzi strains may correlate with infectivity/virulence during the host–parasite interaction.

PARP1-cGAS-NF-κB pathway of proinflammatory macrophage activation by extracellular vesicles released during Trypanosoma cruzi infection and Chagas disease

“…Recent literature documented that this jumping of T. cruzi antigens occurs through the release of membrane vesicles (also called extracellular vesicles). It was shown that T. cruzi sheds compositionally different Ev depending on the developmental stage and virulence of the parasite strain [18,19], Ev shed by infective trypomastigote form of the parasite have high fusogenic potential with the host cell membranes [20], and contact with infective forms of the parasite also stimulated Ca 2+ -dependent shedding of membrane vesicles from THP-1 Mφ [21]. These and other studies did not explore the signaling cascades by which T. cruzi stimulates formation of membrane vesicles within itself or on the host cell membranes, though it was proposed that the host and parasite Ev may maintain cellular activation in CD [7].…”

“…Transfection and dual luciferase assays were conducted by using a Transfection Collection NFκB Transient Pack (79268, BPS Biosciences, San Diego, CA). Briefly, Raw Mφ (30,000 cells/ 100 μL BPS medium) were seeded in 96-well, clear bottom, tissue culture plates, and allowed to adhere for 24 h. For transfection, 1 μL of NFκB reporter (consists NFκB reporter vector + constitutively expressing Renilla luciferase vector) or negative control reporter (non-inducible luciferase vector + Renilla luciferase vector) were diluted in 15 μL of Opti MEM I medium, mixed with 0.35 μL of Lipofectamine 2000, and added to each well [19]. After incubation for 24 h at 37˚C / 5% CO 2 , cells were replenished with fresh BPS medium.…”

Message in a vesicle – trans-kingdom intercommunication at the vector–host interface

“…Trypomastigote vesicles carry molecules involved in nucleic acid binding, heatshock proteins and mucins, as well as parasite surface components (Bayer-Santos et al, 2013;Gonçalves et al, 1991;Nogueira et al, 2015). Because T. cruzi comprises several groups and subgroups, there are strain-dependent differences in vesicle protein content, pathogenesis and immune evasion (Neves et al, 2014;Ribeiro et al, 2018;Wyllie and Ramirez, 2017).…”

Role of Proteomics in the Study of Trypanosoma cruzi Biology

“…cruzi was shown to release vesicles containing a wide range of surface molecules to affect the host immune responses and cell invasion [72]. A proteomics study reported different compositions of extracellular vesicles released by two T. cruzi strains associated with their distinct interaction with host cells [73]. The secretomes of two distinct T. cruzi strains (Y and YuYu), which were previously shown to differentially modulate host immune responses, were compared.…”

“…On the other hand, YuYu strain-derived EVs induced greater levels of infection of LLC-MK2 cells than Y strain-derived EVs. In summary, the quantitative and qualitative differences in EVs and released proteins from distinct T. cruzi strains may be correlated with the parasite capacity to be infective and virulent [73].…”