Chai Yijun, Che Jin, Wang Jinming, Guan Guiquan, Yin Hong
State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, People's Republic of China.
Heilongjiang Provincial Key Laboratory of Zoonosis, College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang, People's Republic of China.
Parasit Vectors. 2025 Jun 10;18(1):221. doi: 10.1186/s13071-025-06834-8.
Extracellular vesicles (EVs) play a pivotal role in host-parasite interactions, particularly in facilitating parasite pathogenesis and immune modulation, and are crucial mediators of intercellular communication. Theileria annulata, an apicomplexan parasite, induces severe alterations in host cells, promoting uncontrolled proliferation, resistance to apoptosis, and immune evasion. Although EVs contribute to these processes, the proteins and small RNA cargo involved in T. annulata infection remain incompletely characterized. In particular, little is known about EV profiles in infections caused by drug-resistant field strains.
In this study, we conducted systematic proteomic and small RNA profiling of EVs derived from naturally occurring buparvaquone-resistant T. annulata (Xinjiang Kashi strain) infected and uninfected bovine sera to investigate infection-induced alterations. Additionally, EVs were isolated from T. annulata-infected bovine immune cells to determine the protein and microRNA (miRNA) compositions of EVs secreted by specific immune cell types. Label-free liquid chromatography-tandem mass spectrometry proteomics and small RNA sequencing were employed to identify EV-associated proteins and miRNAs, followed by functional enrichment analysis to explore key host-parasite regulatory pathways.
Our analysis identified 2580 proteins and 6635 miRNAs in EVs derived from T. annulata-infected bovine serum and immune cell types, many of which are implicated in parasite development, host invasion, and immune modulation. Significant alterations were observed in the EV cargo from infected sera, including enrichment of vesicular proteins and miRNAs associated with immune regulation, metabolic reprogramming, and host-pathogen interactions. Furthermore, functional enrichment analyses highlighted key pathways such as ECM-receptor interactions, oxidative phosphorylation, and proton transport, underscoring the role of EVs in host immune modulation. Supplementary analysis of EVs from infected immune cells provided further insights into the cell type-specific contributions.
This study comprehensively characterized the infection-induced changes in serum-derived EVs associated with a naturally occurring buparvaquone-resistant T. annulata infection. It offers novel insights into how T. annulata exploits EVs to manipulate host responses. The identification of unique EV-associated proteins and miRNAs highlights their potential as biomarkers and therapeutic targets for Theileria infections. These findings contribute to a deeper understanding of host-parasite interactions and lay the foundation for future investigations into EV-mediated pathogenesis and immune evasion.
细胞外囊泡（EVs）在宿主 - 寄生虫相互作用中起关键作用，特别是在促进寄生虫发病机制和免疫调节方面，并且是细胞间通讯的重要介质。环形泰勒虫是一种顶复门寄生虫，可诱导宿主细胞发生严重改变，促进细胞不受控制地增殖、抵抗凋亡以及免疫逃逸。尽管细胞外囊泡参与了这些过程，但与环形泰勒虫感染相关的蛋白质和小RNA货物仍未完全明确。特别是，对于耐药现场菌株引起的感染中的细胞外囊泡特征了解甚少。
在本研究中，我们对来自自然存在的对丁萘醌耐药的环形泰勒虫（新疆喀什株）感染和未感染的牛血清中提取的细胞外囊泡进行了系统的蛋白质组学和小RNA分析，以研究感染引起的变化。此外，从感染环形泰勒虫的牛免疫细胞中分离细胞外囊泡，以确定特定免疫细胞类型分泌的细胞外囊泡的蛋白质和微小RNA（miRNA）组成。采用无标记液相色谱 - 串联质谱蛋白质组学和小RNA测序来鉴定与细胞外囊泡相关的蛋白质和miRNA，随后进行功能富集分析以探索关键的宿主 - 寄生虫调节途径。
我们的分析在来自感染环形泰勒虫的牛血清和免疫细胞类型的细胞外囊泡中鉴定出2580种蛋白质和6635种miRNA，其中许多与寄生虫发育、宿主侵袭和免疫调节有关。在感染血清的细胞外囊泡货物中观察到显著变化，包括与免疫调节、代谢重编程和宿主 - 病原体相互作用相关的囊泡蛋白和miRNA的富集。此外，功能富集分析突出了细胞外基质 - 受体相互作用、氧化磷酸化和质子运输等关键途径，强调了细胞外囊泡在宿主免疫调节中的作用。对感染免疫细胞的细胞外囊泡的补充分析进一步深入了解了细胞类型特异性贡献。
本研究全面表征了与自然存在的对丁萘醌耐药的环形泰勒虫感染相关的血清来源细胞外囊泡中感染诱导的变化。它为环形泰勒虫如何利用细胞外囊泡操纵宿主反应提供了新的见解。独特的与细胞外囊泡相关的蛋白质和miRNA的鉴定突出了它们作为环形泰勒虫感染的生物标志物和治疗靶点的潜力。这些发现有助于更深入地理解宿主 - 寄生虫相互作用，并为未来对细胞外囊泡介导的发病机制和免疫逃逸的研究奠定基础。
