State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, the Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, the Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Vet Microbiol. 2023 Sep;284:109834. doi: 10.1016/j.vetmic.2023.109834. Epub 2023 Jul 21.
Porcine epidemic diarrhea virus (PEDV) is a highly pathogenic enteric coronavirus that causes severe enteritis and lethal watery diarrhea in suckling piglets, leading to tremendous economic losses. Exosomes have been reported to participate in intercellular communication by the transportation of a variety of biological materials, including RNAs, lipids, and proteins. However, PEDV transmission routes have not yet been fully elucidated, and whether exosomes function in PEDV transmission remains unclear. In this study, we extracted and purified exosomes from PEDV-infected Vero cells using a stringent isolation method with a combination of chemical precipitation, ultracentrifugation, and incubation with CD63-labeled magnetic beads. We found that exosomes from PEDV-infected Vero cells contain viral genomic RNA and viral nucleocapsid protein. Furthermore, we demonstrated that the purified exosomes from PEDV-infected cells are capable of transmitting the virus to both PEDV-susceptible and non-susceptible cells. Importantly, exosome-mediated PEDV infection was resistant to neutralization by PEDV-specific neutralizing antibodies that potently neutralized free PEDV. Our study reveals a potential immune evasion mechanism utilized by PEDV and provides new insight into the transmission and infection of this important pathogen.
猪流行性腹泻病毒(PEDV)是一种高致病性肠冠状病毒,可引起仔猪严重的肠炎和致死性水样腹泻,造成巨大的经济损失。外泌体已被报道通过运输包括 RNA、脂质和蛋白质在内的多种生物物质参与细胞间通讯。然而,PEDV 的传播途径尚未完全阐明,外泌体是否在 PEDV 的传播中起作用仍不清楚。在本研究中,我们使用化学沉淀、超速离心和用 CD63 标记的磁珠孵育相结合的严格分离方法,从 PEDV 感染的 Vero 细胞中提取和纯化外泌体。我们发现,来自 PEDV 感染的 Vero 细胞的外泌体含有病毒基因组 RNA 和病毒核衣壳蛋白。此外,我们证明,来自 PEDV 感染细胞的纯化外泌体能够将病毒传播给 PEDV 易感和非易感细胞。重要的是,外泌体介导的 PEDV 感染对能够有效中和游离 PEDV 的 PEDV 特异性中和抗体具有抗性。我们的研究揭示了 PEDV 利用的一种潜在的免疫逃避机制,并为研究这种重要病原体的传播和感染提供了新的见解。