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基于iTRAQ的日本血吸虫童虫和成虫排泄分泌蛋白的比较蛋白质组学分析

iTRAQ-based comparative proteomic analysis of excretory-secretory proteins of schistosomula and adult worms of Schistosoma japonicum.

作者信息

Cao Xiaodan, Fu Zhiqiang, Zhang Min, Han Yanhui, Han Hongxiao, Han Qian, Lu Ke, Hong Yang, Lin Jiaojiao

机构信息

Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai, China.

College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, China.

出版信息

J Proteomics. 2016 Apr 14;138:30-9. doi: 10.1016/j.jprot.2016.02.015. Epub 2016 Feb 23.

Abstract

Schistosomiasis remains a serious public health problem with 200 million people infected and 779 million people at risk worldwide. The schistosomulum and adult worm are two stages of the complex lifecycle of Schistosoma japonicum and excretory/secretory proteins (ESPs) play a major role in host-parasite interactions. In this study, iTRAQ-coupled LC-MS/MS was used to investigate the proteome of ESPs obtained from schistosomula and adult worms of S. japonicum, and 298 differential ESPs were identified. Bioinformatics analysis of differential ESPs in the two developmental stages showed that 161 ESPs upregulated in schistosomula were associated with stress responses, carbohydrate metabolism and protein degradation, whereas ESPs upregulated in adult worms were mainly related to immunoregulation and purine metabolism. Recombinant heat shock protein 70 (HSP70) and thioredoxin peroxidase (TPx), two differential proteins identified in this study, were expressed. Further studies showed that rSjHSP70 and rSjTPx stimulated macrophages expressing high levels of the anti-inflammatory factors TGF-β, IL-10 and Arg-1, and suppressed the expression of the pro-inflammatory cytokines TNF-α, IL-1β, IL-6 and iNOS in LPS-induced macrophages. This study provides new insights into the survival and development of schistosomes in the final host and helps identify vaccine candidates or new diagnostic reagents for schistosomiasis.

摘要

血吸虫病仍然是一个严重的公共卫生问题,全球有2亿人感染,7.79亿人面临风险。童虫和成虫是日本血吸虫复杂生命周期中的两个阶段,排泄/分泌蛋白(ESPs)在宿主-寄生虫相互作用中起主要作用。在本研究中,采用iTRAQ耦合液相色谱-串联质谱法研究日本血吸虫童虫和成虫ESPs的蛋白质组,鉴定出298种差异ESPs。对两个发育阶段差异ESPs的生物信息学分析表明,童虫中上调的161种ESPs与应激反应、碳水化合物代谢和蛋白质降解有关,而成虫中上调的ESPs主要与免疫调节和嘌呤代谢有关。表达了本研究鉴定出的两种差异蛋白,即重组热休克蛋白70(HSP70)和硫氧还蛋白过氧化物酶(TPx)。进一步研究表明,rSjHSP70和rSjTPx刺激巨噬细胞表达高水平的抗炎因子TGF-β、IL-10和Arg-1,并抑制LPS诱导的巨噬细胞中促炎细胞因子TNF-α、IL-1β、IL-6和iNOS的表达。本研究为血吸虫在终末宿主体内的生存和发育提供了新的见解,并有助于确定血吸虫病的候选疫苗或新的诊断试剂。

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