Liu Wenxiao, Hong Jiabing, Duan Jinglong, Jiang Bo, Zhu Runan, Cheng Jing, Wang Ping, Li Yongqing
Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, 100097, Beijing, China.
Research Center for Infectious Diseases in Livestock and Poultry, Beijing Academy of Agricultural and Forestry Sciences, 100097, Beijing, China.
Appl Microbiol Biotechnol. 2023 Jan;107(1):379-390. doi: 10.1007/s00253-022-12308-z. Epub 2022 Dec 2.
Infections caused by bovine herpesvirus 1 (BoHV-1) remain a serious global issue to the health and welfare of the bovine industry. Monitoring of neutralizing antibodies is essential not only for epidemic diagnosis, but also to assess vaccination efficacy. In this study, we generated a neutralizing monoclonal antibody, termed as 3F8, targeting glycoprotein D (gD) of BoHV-1. This monoclonal antibody could neutralize BoHV-1 with a 50% inhibitory concentration (IC) of 37.82 ng/mL. Furthermore, 3F8 could inhibit BoHV-1 infection and cell-to-cell spread at the prebinding stage. A blocking enzyme-linked immunosorbent assay (ELISA) for detecting neutralizing antibodies against BoHV-1 was then developed based on 3F8 and protein gD generated using a baculovirus expression system. The sensitivity and specificity of the test were estimated to be 94.59% and 93.42%, respectively. A significant correlation (R = 0.9583, p < 0.01) was observed between the results obtained with the blocking ELISA and a virus neutralization test, which suggested that the blocking ELISA could detect neutralizing antibodies against BoHV-1. A serological survey was carried out in the dairy farms in Beijing district using 3F8-based blocking ELISA to monitor the annual neutralization antibody against BoHV-1 during 2012-2020. It revealed that the dairy farms in Beijing were at high risk of BoHV-1 infection during 2012-2017 but were protected since 2018 upon implementation of an immunization program. Our results demonstrated that this assay is suitable for BoHV-1 surveillance and vaccination efficacy in cattle as a replacement for the virus neutralization test. KEY POINTS: • Prevention of BoHV-1 infection requires the monitoring of neutralizing antibodies. • A blocking ELISA for the neutralizing antibody was developed based on mAb 3F8 against BoHV-1 gD. • It can replace the labor-intensive and time-consuming viral neutralizing tests.
牛疱疹病毒1型（BoHV-1）引起的感染仍是影响养牛业健康和福利的一个严重全球性问题。监测中和抗体不仅对疫情诊断至关重要，而且对评估疫苗接种效果也很关键。在本研究中，我们制备了一种针对BoHV-1糖蛋白D（gD）的中和单克隆抗体，命名为3F8。该单克隆抗体能够中和BoHV-1，其50%抑制浓度（IC）为37.82 ng/mL。此外，3F8可在结合前阶段抑制BoHV-1感染及细胞间传播。随后基于3F8和利用杆状病毒表达系统产生的gD蛋白，开发了一种用于检测抗BoHV-1中和抗体的阻断酶联免疫吸附测定（ELISA）。该检测方法的敏感性和特异性估计分别为94.59%和93.42%。阻断ELISA检测结果与病毒中和试验结果之间存在显著相关性（R = 0.9583，p < 0.01），这表明阻断ELISA能够检测抗BoHV-1的中和抗体。利用基于3F8的阻断ELISA对北京地区奶牛场进行了血清学调查，以监测2012 - 2020年期间抗BoHV-1的年度中和抗体。结果显示，北京地区的奶牛场在2012 - 2017年期间有较高的BoHV-1感染风险，但自2018年实施免疫计划后得到了保护。我们的结果表明，该检测方法适用于牛群中BoHV-1的监测和疫苗接种效果评估，可替代病毒中和试验。要点：• 预防BoHV-1感染需要监测中和抗体。• 基于抗BoHV-1 gD的单克隆抗体3F8开发了一种中和抗体阻断ELISA。• 它可替代劳动强度大且耗时的病毒中和试验。
