Abstract
ABSTRACTTrypanosoma 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.
Highlights
Trypanosoma cruzi cell surface is covered by a wide range of molecules including members of trans-sialidase (TS/gp85), mucin-like glycoproteins, mucin-associated proteins (MASP) superfamilies, and proteases [1,2,3]
We identified the proteins present in the VF fractions obtained by ultracentrifugation at 100,000 × g, for 16 h, of conditioned medium from trypomastigotes of Y and YuYu strains, which might contain proteins weakly associated with extracellular vesicles (EVs) (Figure 1)
YuYu EVs were enriched with TS family of glycoproteins, mucins, and mucin-associated proteins when compared to those from Y strain
Summary
Trypanosoma cruzi cell surface is covered by a wide range of molecules including members of trans-sialidase (TS/gp85), mucin-like glycoproteins, mucin-associated proteins (MASP) superfamilies, and proteases [1,2,3]. Those molecules are involved in attachment and invasion to the host’s cell by infective forms of the parasite [4,5,6]. Several reports have shown proteomic variations in different T. cruzi stages, including cell derived trypomastigotes and epimastigotes [12,13,14,15,16]. A proteomic analysis of secreted material released by trypomastigotes obtained from Vero cells infected with CL-Brener and VD strains
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