Ramirez M I, Deolindo P, de Messias-Reason I J, Arigi Emma A, Choi H, Almeida I C, Evans-Osses I
Instituto Oswaldo Cruz- Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil.
Universidade Federal de Parana, Curitiba, PR, Brazil.
Cell Microbiol. 2017 Apr;19(4). doi: 10.1111/cmi.12672. Epub 2016 Nov 11.
Extracellular vesicles released from pathogens may alter host cell functions. We previously demonstrated the involvement of host cell-derived microvesicles (MVs) during early interaction between Trypanosoma cruzi metacyclic trypomastigote (META) stage and THP-1 cells. Here, we aim to understand the contribution of different parasite stages and their extracellular vesicles in the interaction with host cells. First, we observed that infective host cell-derived trypomastigote (tissue culture-derived trypomastigote [TCT]), META, and noninfective epimastigote (EPI) stages were able to induce different levels of MV release from THP-1 cells; however, only META and TCT could increase host cell invasion. Fluorescence resonance energy transfer microscopy revealed that THP-1-derived MVs can fuse with parasite-derived MVs. Furthermore, MVs derived from the TCT-THP-1 interaction showed a higher fusogenic capacity than those from META- or EPI-THP-1 interaction. However, a higher presence of proteins from META (25%) than TCT (12%) or EPI (5%) was observed in MVs from parasite-THP-1 interaction, as determined by proteomics. Finally, sera from patients with chronic Chagas disease at the indeterminate or cardiac phase differentially recognized antigens in THP-1-derived MVs resulting only from interaction with infective stages. The understanding of intracellular trafficking and the effect of MVs modulating the immune system may provide important clues about Chagas disease pathophysiology.
病原体释放的细胞外囊泡可能会改变宿主细胞的功能。我们之前证明了宿主细胞衍生的微囊泡（MVs）在克氏锥虫循环后期锥鞭毛体（META）阶段与THP-1细胞早期相互作用过程中的作用。在此，我们旨在了解不同寄生虫阶段及其细胞外囊泡在与宿主细胞相互作用中的贡献。首先，我们观察到感染性的宿主细胞衍生锥鞭毛体（组织培养衍生锥鞭毛体 [TCT]）、META和非感染性的上鞭毛体（EPI）阶段能够诱导THP-1细胞释放不同水平的MVs；然而，只有META和TCT能够增加宿主细胞的侵袭。荧光共振能量转移显微镜显示，THP-1衍生的MVs可以与寄生虫衍生的MVs融合。此外，TCT-THP-1相互作用衍生的MVs比META-或EPI-THP-1相互作用衍生的MVs具有更高的融合能力。然而，通过蛋白质组学测定，在寄生虫-THP-1相互作用衍生的MVs中，观察到来自META的蛋白质（25%）比来自TCT（12%）或EPI（5%）的蛋白质含量更高。最后，处于不确定期或心脏期的慢性恰加斯病患者的血清在THP-1衍生的MVs中差异识别仅由与感染性阶段相互作用产生的抗原。对细胞内运输以及MVs调节免疫系统的作用的理解可能为恰加斯病的病理生理学提供重要线索。
