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日本血吸虫质膜修复相关基因肌铁蛋白的特性分析

Characterization Analysis of Schistosoma japonicum Plasma Membrane Repair Relative Gene Myoferlin.

作者信息

Xiong Yanian, Zhang Ming, Hong Yang, Wei Meimei, Ai Dezhou, Meng Peipei, Han Yanhui, Fu Zhiqiang, Shi Yaojun, Yang Jianmei, Lin Jiaojiao

机构信息

National Laboratory of Animal Schistosomiasis Control/Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.

出版信息

PLoS One. 2013 Jun 18;8(6):e66396. doi: 10.1371/journal.pone.0066396. Print 2013.

Abstract

Myoferlin is a member of the ferlin family of proteins, which are involved in plasma membrane repair, and has been identified as one of the tegument proteins of Schistosoma japonicum. The tegument proteins are potential candidates for vaccines and new drug targets. In this study, myoferlin of S. japonicum (SjMF) was cloned, expressed and characterized, the potential of SjMF recombinant protein (rSjMF) as a vaccine candidate was evaluated, and the effect of praziquantel on SjMF was detected by Real-time PCR. Immunofluorescence showed that this protein was mainly distributed on the surface of worms at different stages. Sequence analysis revealed that the SjMF open reading frame was conserved at all stages of the S. japonicum life cycle. And SjMF transcription was upregulated in 42-day-old worms, and was significantly higher in female worms. Western blotting revealed that rSjMF showed strong immunogenicity. The cytokine profile and IgG isotype analysis demonstrated that rSjMF plus ISA206 immunization induced a mixed T helper (Th)1/Th2 response. Purified rSjMF emulsified with ISA206 adjuvant significantly reduced worm burden from 21.8% to 23.21% and liver egg number from42.58% to 28.35%. Besides, SjMF transcription was downregulated when worms were exposed to low-dose praziquantel (PZQ) and upregulated when PZQ was degraded, accompanied by recovery of damaged tegument. When worms were exposed to high-dose PZQ, SjMF transcription was downregulated all the time and the damaged tegument did not recover. These findings indicated that SjMF is a potential vaccine against S. japonicum and provides the basis for further investigations into the biological function of SjMF.

摘要

肌铁蛋白是参与质膜修复的铁蛋白家族蛋白质成员之一,已被鉴定为日本血吸虫的皮层蛋白之一。皮层蛋白是疫苗和新药物靶点的潜在候选物。在本研究中,克隆、表达并鉴定了日本血吸虫的肌铁蛋白(SjMF),评估了SjMF重组蛋白(rSjMF)作为候选疫苗的潜力,并通过实时PCR检测了吡喹酮对SjMF的影响。免疫荧光显示该蛋白主要分布在不同阶段虫体的表面。序列分析表明,SjMF开放阅读框在日本血吸虫生命周期的各个阶段均保守。并且SjMF转录在42日龄虫体中上调,在雌虫中显著更高。蛋白质印迹分析显示rSjMF具有很强的免疫原性。细胞因子谱和IgG亚型分析表明,rSjMF加ISA206免疫诱导了混合的辅助性T细胞(Th)1/Th2反应。用ISA206佐剂乳化的纯化rSjMF显著降低了虫负荷,从21.8%降至23.21%,肝脏虫卵数从42.58%降至28.35%。此外,当虫体暴露于低剂量吡喹酮(PZQ)时,SjMF转录下调,当PZQ降解时上调,同时受损皮层恢复。当虫体暴露于高剂量PZQ时,SjMF转录一直下调,受损皮层未恢复。这些发现表明SjMF是一种潜在的抗日本血吸虫疫苗,并为进一步研究SjMF的生物学功能提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aea6/3688920/e7f348f2069b/pone.0066396.g001.jpg

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