Ge Guiyang, Li Dongli, Ling Qian, Xu Lihui, Ata Emad Beshir, Wang Xiaolin, Li Keyan, Hao Wen, Gong Qinglong, Li Jianming, Shi Kun, Leng Xue, Du Rui
College of Animal Science and Technology, Jilin Agricultural University, Changchun, China.
Wengniute Banner Agriculture and Animal Husbandry Bureau, Chifeng, China.
Front Microbiol. 2024 Dec 3;15:1483527. doi: 10.3389/fmicb.2024.1483527. eCollection 2024.
(IBR), characterized by acute respiratory lesions in cattle, is a major infectious disease caused by (BoAHV-1). Control of this disease is primarily depending on vaccination. Madin-Darby bovine kidney cells (MDBK) being the main host cells and the important production platform for IBR vaccines. However, innate immune genes inhibit viral replication. Accordingly, the aim of this study was developing of gene deleted MDBK cells to facilitate the production of high-titer vaccines. The CRISPR/Cas9 technology was used to knock out the gene in MDBK cells and the impact on virus replication was examined using virus growth curves, CCK-8 assays, cell scratch assays, and qPCR. The knockout of the IRF7 gene in MDBK cells led to an increased replication capacity of IBRV and a significant reduction in type I interferons expression, specifically IFN-α and IFN-β. This indicates that MDBK cell lines can effectively result in production of IBRV with high-titer, which will enhance the development of inactivated or attenuated vaccines.
传染性牛鼻气管炎（IBR）以牛的急性呼吸道病变为特征，是由牛疱疹病毒1型（BoAHV-1）引起的一种主要传染病。该病的防控主要依赖于疫苗接种。马德堡-达比牛肾细胞（MDBK）是IBR疫苗的主要宿主细胞和重要生产平台。然而，天然免疫基因会抑制病毒复制。因此，本研究的目的是构建IRF7基因缺失的MDBK细胞，以促进高滴度疫苗的生产。利用CRISPR/Cas9技术敲除MDBK细胞中的IRF7基因，并通过病毒生长曲线、CCK-8检测、细胞划痕试验和qPCR检测对病毒复制的影响。MDBK细胞中IRF7基因的敲除导致IBR病毒复制能力增强，I型干扰素尤其是IFN-α和IFN-β的表达显著降低。这表明，IRF7基因缺失的MDBK细胞系能够有效产生高滴度的IBR病毒，这将促进灭活疫苗或减毒活疫苗的研发。
