Wang Qi, Chen Tuanjie, Feng Mengru, Zheng Mei, Gao Feixia, Qiu Chenchen, Luo Jian, Li Xiuling
Department of Virus and Vaccine, Shanghai Institute of Biological Products, Shanghai 200052, China.
Viruses. 2025 Aug 8;17(8):1097. doi: 10.3390/v17081097.
Cell culture-based influenza vaccines exhibit comparable safety and immunogenicity to traditional egg-based vaccines. However, improving viral yield remains a key challenge in optimizing cell culture-based production systems. Madin-Darby canine kidney (MDCK) cells, the predominant cell line for influenza vaccine production, inherently activate interferon (IFN)-mediated antiviral defenses that restrict viral replication. To overcome this limitation, we employed CRISPR/Cas9 gene-editing technology to generate an IFN alpha/beta receptor subunit 1 (IFNAR1)-knockout (KO) adherent MDCK cell line. Viral titer analysis demonstrated significant enhancements in the yield of multiple vaccine strains (H1N1, H3N2, and type B) in IFNAR1-KO cells compared to wild-type (WT) cells. Transcriptomic profiling revealed marked downregulation of key interferon-stimulated genes (ISGs)-including , , and -within the IFNAR1-KO cells, indicating a persistent suppression of antiviral responses that established a more permissive microenvironment for influenza virus replication. Collectively, the engineered IFNAR1-KO cell line provides a valuable tool for influenza virus research and a promising strategy for optimizing large-scale MDCK cell cultures to enhance vaccine production efficiency.
基于细胞培养的流感疫苗与传统的基于鸡蛋的疫苗相比，具有相当的安全性和免疫原性。然而，提高病毒产量仍然是优化基于细胞培养的生产系统的关键挑战。流感疫苗生产的主要细胞系——犬肾传代细胞（MDCK）细胞，固有地激活干扰素（IFN）介导的抗病毒防御，从而限制病毒复制。为了克服这一限制，我们采用CRISPR/Cas9基因编辑技术生成了一种干扰素α/β受体亚基1（IFNAR1）基因敲除（KO）的贴壁MDCK细胞系。病毒滴度分析表明，与野生型（WT）细胞相比，IFNAR1-KO细胞中多种疫苗株（H1N1、H3N2和B型）的产量有显著提高。转录组分析显示，IFNAR1-KO细胞内关键的干扰素刺激基因（ISGs）——包括 、 和 显著下调，表明抗病毒反应持续受到抑制，从而为流感病毒复制建立了一个更宽松的微环境。总的来说，工程化的IFNAR1-KO细胞系为流感病毒研究提供了一个有价值的工具，也是优化大规模MDCK细胞培养以提高疫苗生产效率的一个有前景的策略。
