Isolation of exosomes from Trichinella spiralis muscle larvae and identification of small RNAs
GAO Xin, YANG Yong, LIU Lei, LIU Xiao-lei, LIU Ming-yuan, BAI Xue
Institute of Zoonosis, Key Laboratory of Zoonosis Research, Ministry of Education, College of Veterinary Medicine, Jilin University, Changchun 130062, China
Abstract:Trichinella spiralis (T. spiralis) is an important food-born parasite that can regulate the host immune response to achieve long-term parasitic. Recently, it has been found that exosomes derived from parasites can not only transmit virulence factors between parasites, but also regulate the immune response of hosts. We purified and identified T. spiralis muscle larvae exosomes, providing a new direction for the research on the immune regulation ability of T. spiralis. In this experiment, exosomes of T. spiralis muscle larvae were extracted by ultracentrifugation and were identified by transmission electron microscope, nanoparticle tracking analysis, western blotting and small RNA sequencing. T. spiralis muscle larvae exosomes were membrane-structure vesicles with a diameter of 80-200 nm and expressed exosomes-specific markers CD63 and Enolase, containing 1266 known miRNAs. Our study indicated that the exosomes from Trichinella spiralis muscle larvae were successfully extracted and were comfirmed by the morphology, particle size and markers analyze. Meanwhile,small RNA libraries of exosome were constructure to indentify a variety of functional small RNAs. These findings provide data for further study the application of small RNAs in exosomes.
高欣, 杨勇, 刘蕾, 刘晓雷, 刘明远, 白雪. 旋毛虫肌幼虫期外泌体的分离和小RNA鉴定[J]. 中国人兽共患病学报, 2020, 36(4): 261-266.
GAO Xin, YANG Yong, LIU Lei, LIU Xiao-lei, LIU Ming-yuan, BAI Xue. Isolation of exosomes from Trichinella spiralis muscle larvae and identification of small RNAs. Chinese Journal of Zoonoses, 2020, 36(4): 261-266.
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