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鼠细胞外囊泡的特性与弓形虫感染。

Characterization of murine extracellular vesicles and Toxoplasma gondii infection.

机构信息

Centro de Parasitologia e Micologia, Instituto Adolfo Lutz, Sao Paulo, Brazil.

Nucleo de Microscopia Eletrônica, Instituto Adolfo Lutz, Sao Paulo, Brazil.

出版信息

Parasite Immunol. 2021 Sep;43(9):e12869. doi: 10.1111/pim.12869. Epub 2021 Jun 7.

Abstract

This study characterized extracellular vesicles (EVs) of sera from mice infected with Toxoplasma gondii or immunized with EVs derived T gondii. EVs were purified of sera from four groups (5 A/Sn mice/group). EV-IM: Mice immunized with T gondii-released EVs; ACT: mice in acute infection; CHR: mice in chronic infection; and NI: normal mice. EVs were purified by ultracentrifugation. Concentration of serum-derived EVs from NI group was smaller than EV-IM, ACT and CHR groups. Most of the EVs from ACT and CHR groups were microvesicles, and they were bigger than the NI group. The same results were shown by Transmission Electron Microscopy. The presence of exosomes was shown in immunoblotting by tetraspanin (CD63 and CD9) evidence. Splenocytes of EV-IM, CHR and NI groups were stimulated with T. gondii derived EVs. EV-IM and CHR groups up-expressed IFN-γ; TNF-α and IL-17, when compared with the NI group. IL-10 was up-expressed only in the EV-IM group. EV-IM, ACT and CHR groups expressed more miR-155-5p, miR-29c-3p and miR-125b-5p than the NI group. Host-T gondii interaction can occur, also, via EVs. miRNAs participate in the modulation of cellular immune response against T gondii. These data give subsidies to propose the differentiation between infect or noninfect hosts by concentration of EVs.

摘要

本研究对感染弓形虫的小鼠或用弓形虫来源的 EV 免疫的小鼠血清中的细胞外囊泡(EVs)进行了特征分析。从 4 组(每组 5 只 A/Sn 小鼠)血清中纯化 EV:EV-IM:用弓形虫释放的 EV 免疫的小鼠;ACT:急性感染小鼠;CHR:慢性感染小鼠;NI:正常小鼠。EV 通过超速离心进行纯化。NI 组血清来源的 EV 浓度小于 EV-IM、ACT 和 CHR 组。ACT 和 CHR 组的大多数 EV 为微囊泡,且大于 NI 组。透射电子显微镜显示了相同的结果。免疫印迹显示四跨膜蛋白(CD63 和 CD9)存在外泌体的证据。EV-IM、CHR 和 NI 组的脾细胞用弓形虫来源的 EV 刺激。与 NI 组相比,EV-IM 和 CHR 组 IFN-γ、TNF-α 和 IL-17 表达上调;仅在 EV-IM 组中表达 IL-10。与 NI 组相比,EV-IM、ACT 和 CHR 组表达更多的 miR-155-5p、miR-29c-3p 和 miR-125b-5p。宿主-弓形虫相互作用也可以通过 EV 发生。miRNAs 参与调节针对弓形虫的细胞免疫反应。这些数据支持通过 EV 浓度来区分感染或非感染宿主的假说。

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