He Xinglin, Li Pengfei, Cao Hua, Zhang Xiaoling, Zhang Mengjia, Yu Xuexiang, Sun Yumei, Ghonaim Ahmed H, Ma Hailong, Li Yongtao, Shi Kaizhi, Zhu Hongmei, He Qigai, Li Wentao
National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The Cooperative Innovation Center for Sustainable Pig Production, Wuhan, 430070, China; Hubei Hongshan Laboratory, Wuhan, 430070, China.
College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, China.
Antiviral Res. 2025 Jan;233:106058. doi: 10.1016/j.antiviral.2024.106058. Epub 2024 Dec 11.
African Swine Fever (ASF) is a highly lethal and contagious disease in pigs caused by African Swine Fever Virus (ASFV), which primarily infects domestic pigs and wild boars, with a mortality rate of up to 100%. Currently, there are no commercially available vaccines or drugs that are both safe and effective against ASFV. The ASFV 0428C strain was continuously passaged in Vero cells, and the adapted ASFV demonstrated efficient replication in Vero cells. The adapted ASFV was used as the parental virus, and an expression cassette encoding a dual reporter gene for firefly luciferase (Fluc) and enhanced green fluorescent protein (eGFP) was inserted into the ASFV genome using CRISPR/Cas9 gene editing technology to construct a recombinant ASFV variant (rASFV-FLuc-eGFP). rASFV-Fluc-eGFP was genetically stable, effectively infected porcine alveolar macrophages (PAM) and Vero cells, and expressed Fluc and eGFP concurrently. This study provides a tool for investigating the infection and pathogenic mechanisms of ASFV, as well as for screening essential host genes and antiviral drugs. Additionally, a high-throughput screening model of antiviral drugs was established based on rASFV-FLuc-eGFP in passaged cells, 218 compounds from the FDA-approved compound library were screened, and 5 candidate compounds with significant inhibitory effects in Vero cells were identified. The inhibitory effects on ASFV were further validated in both Vero and PAM cells, resulting in the identification of Salvianolic acid C (SAC), which demonstrated inhibitory effects and safety in both cell types. SAC is a candidate drug for the prevention and control of ASFV and shows promising application prospects.
非洲猪瘟（ASF）是由非洲猪瘟病毒（ASFV）引起的猪的一种高致死性和传染性疾病，主要感染家猪和野猪，死亡率高达100%。目前，尚无既安全又有效的针对ASFV的商业可用疫苗或药物。ASFV 0428C毒株在Vero细胞中连续传代，适应后的ASFV在Vero细胞中表现出高效复制。将适应后的ASFV作为亲本病毒，利用CRISPR/Cas9基因编辑技术将编码萤火虫荧光素酶（Fluc）和增强型绿色荧光蛋白（eGFP）双报告基因的表达盒插入ASFV基因组，构建重组ASFV变异株（rASFV-FLuc-eGFP）。rASFV-FLuc-eGFP遗传稳定，能有效感染猪肺泡巨噬细胞（PAM）和Vero细胞，并同时表达Fluc和eGFP。本研究为研究ASFV的感染和致病机制、筛选关键宿主基因和抗病毒药物提供了一种工具。此外，基于传代细胞中的rASFV-FLuc-eGFP建立了抗病毒药物高通量筛选模型，对FDA批准的化合物库中的218种化合物进行了筛选，鉴定出5种在Vero细胞中具有显著抑制作用的候选化合物。在Vero细胞和PAM细胞中进一步验证了其对ASFV的抑制作用，鉴定出丹酚酸C（SAC），其在两种细胞类型中均表现出抑制作用且具有安全性。SAC是预防和控制ASFV的候选药物，具有广阔的应用前景。
