利用生物发光成像技术研发一种重组鳗疱疹病毒1型候选疫苗，该疫苗在体外可正常复制，但在体内感染失败。

Development Using Bioluminescence Imaging of a Recombinant Anguillid Herpesvirus 1 Vaccine Candidate Associated with Normal Replication In Vitro but Abortive Infection In Vivo.

作者信息

Zhang Haiyan, Sridhar Arun, Delrez Natacha, He Bo, Fourny Sophie, Gao Yuan, Donohoe Owen, Vanderplasschen Alain F C

机构信息

Immunology-Vaccinology, Department of Infectious and Parasitic Diseases, Fundamental and Applied Research for Animals & Health (FARAH), Faculty of Veterinary Medicine, University of Liège, B-4000 Liège, Belgium.

Bioscience Research Institute, Technological University of the Shannon, N37 HD68 Athlone, Westmeath, Ireland.

出版信息

Vaccines (Basel). 2024 Dec 17;12(12):1423. doi: 10.3390/vaccines12121423.

DOI:10.3390/vaccines12121423

PMID:39772083

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11728778/

Abstract

BACKGROUND/OBJECTIVES: Anguillid herpesvirus 1 (AngHV-1) (recently renamed Cyvirus anguillidallo 1) is the etiologic agent of a lethal disease that affects several eel species. It is thought to be one of the main infectious agents causing a population decline in wild eels and economic loss within the eel aquaculture sector. To date, no vaccines are available against AngHV-1. Recently, we developed a safe and efficacious live attenuated recombinant vaccine against Cyprinid herpesvirus 3 (CyHV-3). This CyHV-3 recombinant vaccine encodes a deletion of ORF57. Orthologues of CyHV-3 ORF57 exist in Cyprinid herpesvirus 2 (CyHV-2, ORF57) and AngHV-1 (ORF35).

METHODS

In the present study, using recombinant strains and bioluminescent in vivo imaging, we investigated the effect of AngHV-1 ORF35 deletion on virus replication in vitro, virulence in vivo, and the potential of an AngHV-1 ORF35-deleted recombinant as a vaccine candidate for the mass vaccination of eels by immersion. With this goal in mind, we produced ORF35-deleted recombinants using two parental strains: a UK strain and a recombinant derived from the former strain by insertion of a Luciferase-GFP reporter cassette into a non-coding intergenic region.

RESULTS

Analyses of ORF35-deleted recombinants led to the following observations: (i) AngHV-1 ORF35 is not essential for viral growth in cell culture, and its deletion does not affect the production of extracellular virions despite reducing the size of viral plaque. (ii) In contrast to what has been observed for CyHV-3 ORF57 and CyHV-2 ORF57, in vivo bioluminescent analyses revealed that AngHV-1 ORF35 is an essential virulence factor and that its deletion led to abortive infection in vivo. (iii) Inoculation of the AngHV-1 ORF35-deleted recombinant by immersion induced a protective immune response against a wild-type challenge. This protection was shown to be dose-dependent and to rely on the infectivity of AngHV-1 ORF35-deleted virions.

CONCLUSIONS

This study suggests that the AngHV-1 ORF35 protein has singular properties compared to its orthologues encoded by CyHV-2 and CyHV-3. It also supports the potential of AngHV-1 ORF35-deleted recombinants for the mass vaccination of eels by immersion.

摘要

背景/目的：鳗鲡疱疹病毒1（AngHV-1）（最近重新命名为Cyvirus anguillidallo 1）是一种致死性疾病的病原体，可感染多种鳗鱼品种。它被认为是导致野生鳗鱼种群数量下降和鳗鱼养殖行业经济损失的主要感染因子之一。迄今为止，尚无针对AngHV-1的疫苗。最近，我们开发了一种安全有效的针对鲤疱疹病毒3（CyHV-3）的减毒活重组疫苗。这种CyHV-3重组疫苗编码缺失ORF57。CyHV-3 ORF57的直系同源物存在于鲤疱疹病毒2（CyHV-2，ORF57）和AngHV-1（ORF35）中。

方法

在本研究中，我们使用重组菌株和体内生物发光成像技术，研究了缺失AngHV-1 ORF35对病毒体外复制、体内毒力的影响，以及缺失AngHV-1 ORF35的重组体作为鳗鱼浸泡式大规模疫苗候选物的潜力。出于这个目的，我们使用两种亲本菌株制备了缺失ORF35的重组体：一种英国菌株和一种通过将荧光素酶-绿色荧光蛋白报告基因盒插入非编码基因间隔区而从前者衍生而来的重组体。

结果

对缺失ORF35的重组体的分析得出以下观察结果：（i）AngHV-1 ORF35对于细胞培养中的病毒生长不是必需的，其缺失不影响细胞外病毒粒子的产生，尽管会减小病毒蚀斑的大小。（ii）与在CyHV-3 ORF57和CyHV-2 ORF57中观察到的情况相反，体内生物发光分析表明AngHV-1 ORF35是一种必需的毒力因子，其缺失导致体内感染失败。（iii）通过浸泡接种缺失AngHV-1 ORF35的重组体可诱导针对野生型攻击的保护性免疫反应。这种保护作用显示出剂量依赖性，并依赖于缺失AngHV-1 ORF35的病毒粒子的感染性。