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日本血吸虫中间宿主钉螺中14 kDa硫氧还蛋白相关蛋白的鉴定及功能特性分析

Identification and functional characterization of thioredoxin-related protein of 14 kDa in Oncomelania hupensis, the intermediate host of Schistosoma japonicum.

作者信息

Cao Yunchao, Huang Shuaiqin, Peng Wuxian, Lu Mingke, Peng Wenfeng, Lin Jiaojiao, Tang Chongti, Tang Liang

机构信息

State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, 361102, Fujian, China; Parasitology Research Laboratory, School of Life Sciences, Xiamen University, Xiamen, 361102, Fujian, China.

State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, 361102, Fujian, China; Parasitology Research Laboratory, School of Life Sciences, Xiamen University, Xiamen, 361102, Fujian, China.

出版信息

Mol Biochem Parasitol. 2018 Oct;225:38-46. doi: 10.1016/j.molbiopara.2018.08.009. Epub 2018 Aug 31.

Abstract

Oncomelania hupensis is the unique intermediate host of the blood fluke Schistosoma japonicum, which causes schistosomiasis. In snails, highly toxic reactive oxygen species (ROS) can be continually generated by hemocytes in response to foreign particles or pathogens, and may be involved in damaging and eliminating digenean larvae. Thioredoxin-related protein of 14 kDa (TRP14) is a member of the Trx superfamily, and plays an important role in the scavenging of ROS. This study was designed to identify and characterize TRP14 from O. hupensis (OhTRP14), and investigate the involvement of OhTRP14 in the scavenging of ROS in snail host immune response to the parasite S. japonicum. Here we expressed and purified the recombinant OhTRP14 and its mutant, and rOhTRP14 displayed oxidoreductase activity dependent on the CPDC motif. OhTRP14 protein was ubiquitously present in all the tested snail tissues, and especially immunolocalized in the cytoplasm of immune cell types (hemocytes). Both the expression of OhTRP14 and ROS level increased significantly in snails following challenge with S. japonicum. The dsRNA-mediated knockdown of OhTRP14 was successfully conducted by oral feeding, and ROS production was increased by OhTRP14 knockdown, implying that OhTRP14 was involved in the scavenging of ROS in O. hupensis circulating hemocytes. Therefore, we conclude that OhTRP14 may be involved in the scavenging of ROS in snail host immune response to the parasite S. japonicum. The results expand our understanding of the interaction between this parasite and host, and lay a foundation for the establishment of Oncomelania-schistosome infection models.

摘要

钉螺是日本血吸虫的唯一中间宿主,日本血吸虫可导致血吸虫病。在钉螺中,血细胞会对外来颗粒或病原体产生反应,持续产生高毒性活性氧(ROS),ROS可能参与破坏和清除复殖吸虫幼虫。14 kDa硫氧还蛋白相关蛋白(TRP14)是硫氧还蛋白超家族的成员,在清除ROS中起重要作用。本研究旨在鉴定和表征钉螺中的TRP14(OhTRP14),并研究OhTRP14在钉螺宿主对日本血吸虫免疫反应中清除ROS的作用。在此,我们表达并纯化了重组OhTRP14及其突变体,rOhTRP14表现出依赖CPDC基序的氧化还原酶活性。OhTRP14蛋白普遍存在于所有测试的钉螺组织中,尤其免疫定位在免疫细胞类型(血细胞)的细胞质中。用日本血吸虫攻击后,钉螺体内OhTRP14的表达和ROS水平均显著增加。通过口服成功进行了dsRNA介导的OhTRP14敲低,OhTRP14敲低后ROS产量增加,这意味着OhTRP14参与了钉螺循环血细胞中ROS的清除。因此,我们得出结论,OhTRP14可能参与了钉螺宿主对日本血吸虫免疫反应中ROS的清除。这些结果扩展了我们对这种寄生虫与宿主之间相互作用的理解,并为建立钉螺-血吸虫感染模型奠定了基础。

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