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用 ELPylated 鸭干扰素-α保护雏鸭免受鸭肝炎 A 病毒感染。

Protection of Ducklings from Duck Hepatitis A Virus Infection with ELPylated Duck Interferon-α.

机构信息

Department of Veterinary Medicine, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, China.

College of Veterinary Medicine, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.

出版信息

Viruses. 2022 Mar 18;14(3):633. doi: 10.3390/v14030633.

Abstract

Duck viral hepatitis type I (DVH I) is a lethal disease in ducklings caused by duck hepatitis A virus (DHAV). Although the commercial vaccine is available for vaccination of one-day-old ducklings or breeder ducks, the disease is still prevalent due to the delayed immune response in ducklings and variable maternal antibody levels in breeder duck flocks. To explore the feasibility of duck interferon-α (DuIFN-α) for control of DVH I, DuIFN-α was expressed as an elastin-like polypeptide (ELP) fusion protein (ELP-DuIFN-α) in E. coli and purified by inverse phase transition cycling (ITC). After detection of its cytotoxicity, bioactivity, plasma stability and serum half-life, the protective efficacy of ELP-DuIFN-α against DHAV-1 infection of embryos or ducklings was evaluated using different treatment routes at different infection times. The results show that ELP-DuIFN-α was correctly expressed and purified to more than 90% purity after two cycles of ITC. The purified fusion protein had a specific anti-DHAV-1 activity of 6.0 × 104 IU/mg protein, significantly extended plasma stability and serum half-life without overt cytotoxicity. After allantoic injection with ELP-DuIFN-α pre-infection, co-infection or post-infection with DHAV-1, 5/5, 5/5 or 4/5 embryos survived from the virus challenge. After intramuscular injection or oral administration with ELP-DuIFN-α, 3/5 or 4/5 ducklings survived from co-infection with DHAV-1. After oral administration with ELP-DuIFN-α pre-infection, co-infection or post-infection with DHAV-1, 3/5, 4/5 or 4/5 ducklings survived from the virus challenge, and the relative transcription levels of interferon-stimulated genes were significantly higher than the normal control group and virus challenge control group (p < 0.01). These experimental data suggest that ELP-DuIFN-α can be used as a long-lasting anti-DHAV-1 reagent.

摘要

鸭病毒性肝炎 I 型(DVH I)是一种由鸭肝炎 A 病毒(DHAV)引起的雏鸭致死性疾病。虽然已有针对 1 日龄雏鸭或种鸭的商品化疫苗,但由于雏鸭的免疫反应延迟和种鸭群中母源抗体水平的变化,该病仍很流行。为了探索鸭干扰素-α(DuIFN-α)控制 DVH I 的可行性,本研究在大肠杆菌中表达了作为弹性蛋白样多肽(ELP)融合蛋白(ELP-DuIFN-α),并通过反相相变循环(ITC)进行纯化。在检测其细胞毒性、生物活性、血浆稳定性和血清半衰期后,采用不同的感染时间和处理途径,评估了 ELP-DuIFN-α 对 DHAV-1 感染胚胎或雏鸭的保护效果。结果表明,经过两轮 ITC 后,ELP-DuIFN-α 得到了正确表达和纯化,纯度超过 90%。纯化的融合蛋白具有特异的抗 DHAV-1 活性,为 6.0×104 IU/mg 蛋白,显著延长了血浆稳定性和血清半衰期,且无明显的细胞毒性。在 DHAV-1 前感染、共感染或后感染时进行羊水内注射 ELP-DuIFN-α,5/5、5/5 或 4/5 个胚胎从病毒攻击中存活下来。通过肌肉内注射或口服 ELP-DuIFN-α,3/5 或 4/5 只雏鸭从 DHAV-1 共感染中存活下来。在 DHAV-1 前感染、共感染或后感染时进行口服 ELP-DuIFN-α,3/5、4/5 或 4/5 只雏鸭从病毒攻击中存活下来,干扰素刺激基因的相对转录水平明显高于正常对照组和病毒攻击对照组(p<0.01)。这些实验数据表明,ELP-DuIFN-α可用作一种长效抗 DHAV-1 试剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/567f/8949671/957f105dddb2/viruses-14-00633-g001.jpg

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