A novel high-throughput screen identifies phenazine-1-carboxylic acid as an inhibitor of African swine fever virus replication in primary porcine alveolar macrophages
Abstract African swine fever (ASF), caused by African swine fever virus (ASFV), has resulted in significant economic impacts on the global swine industry. Currently, there is no safe and effective commercial vaccine available for ASFV. Thus, the development of effective and readily available therape...
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BMC
2025-02-01
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| Series: | Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s13567-025-01467-2 |
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| author | Jing Lan Rui Luo Di Liu Changxing Qi Xin Song Zhanhao Lu Ruojia Huang Yuying Yang Yuan Sun Yonghui Zhang Tao Wang Hua-Ji Qiu |
| author_facet | Jing Lan Rui Luo Di Liu Changxing Qi Xin Song Zhanhao Lu Ruojia Huang Yuying Yang Yuan Sun Yonghui Zhang Tao Wang Hua-Ji Qiu |
| author_sort | Jing Lan |
| collection | DOAJ |
| description | Abstract African swine fever (ASF), caused by African swine fever virus (ASFV), has resulted in significant economic impacts on the global swine industry. Currently, there is no safe and effective commercial vaccine available for ASFV. Thus, the development of effective and readily available therapeutics for ASF is urgently needed. To conduct high-throughput screening (HTS) for anti-ASFV drugs, we initially developed a recombinant dual-reporter virus (rASFV-Gluc/EGFP) using the virulent strain ASFV HLJ/18 (ASFV-WT). The enhanced green fluorescent protein (EGFP)- and Gaussia luciferase (Gluc)-encoding genes were incorporated downstream of the ASFV MGF300-4L gene without disrupting viral genes. The growth kinetics, hemadsorption, and transmission electron microscopy analysis of rASFV-Gluc/EGFP in primary porcine alveolar macrophages (PAMs) revealed that rASFV-Gluc/EGFP exhibits similar biological characteristics to ASFV-WT. Furthermore, analysis of Gluc activities, fluorescence, and next-generation sequencing indicated that rASFV-Gluc/EGFP maintains good genetic stability after 20 consecutive passages in PAMs. Using the HTS platform established with rASFV-Gluc/EGFP, we screened and identified phenazine-1-carboxylic acid (PCA) as an effective inhibitor of ASFV replication from 246 small molecule compounds in PAMs. Importantly, PCA was found to reduce ASFV replication by as much as 100-fold at a concentration of 25 μM. Overall, this study suggests that rASFV-Gluc/EGFP is suitable for rapid screening of anti-ASFV drugs. Importantly, we showed that PCA has significant anti-ASFV activity in PAMs. |
| format | Article |
| id | doaj-art-4f235baddf044657891d74141cc485bc |
| institution | Kabale University |
| issn | 1297-9716 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Veterinary Research |
| spelling | doaj-art-4f235baddf044657891d74141cc485bc2025-02-09T12:50:11ZengBMCVeterinary Research1297-97162025-02-0156111510.1186/s13567-025-01467-2A novel high-throughput screen identifies phenazine-1-carboxylic acid as an inhibitor of African swine fever virus replication in primary porcine alveolar macrophagesJing Lan0Rui Luo1Di Liu2Changxing Qi3Xin Song4Zhanhao Lu5Ruojia Huang6Yuying Yang7Yuan Sun8Yonghui Zhang9Tao Wang10Hua-Ji Qiu11State Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteState Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteState Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteHubei Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation, Tongji Medical College, Huazhong University of Science and TechnologyState Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteState Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteState Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteCollege of Animal Science and Technology, Yangtze UniversityState Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteHubei Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation, Tongji Medical College, Huazhong University of Science and TechnologyState Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteState Key Laboratory for Animal Disease Control and Prevention, National African Swine Fever Para-Reference Laboratory, National High Containment Facilities for Animal Disease Control and Prevention, Chinese Academy of Agricultural Sciences, Harbin Veterinary Research InstituteAbstract African swine fever (ASF), caused by African swine fever virus (ASFV), has resulted in significant economic impacts on the global swine industry. Currently, there is no safe and effective commercial vaccine available for ASFV. Thus, the development of effective and readily available therapeutics for ASF is urgently needed. To conduct high-throughput screening (HTS) for anti-ASFV drugs, we initially developed a recombinant dual-reporter virus (rASFV-Gluc/EGFP) using the virulent strain ASFV HLJ/18 (ASFV-WT). The enhanced green fluorescent protein (EGFP)- and Gaussia luciferase (Gluc)-encoding genes were incorporated downstream of the ASFV MGF300-4L gene without disrupting viral genes. The growth kinetics, hemadsorption, and transmission electron microscopy analysis of rASFV-Gluc/EGFP in primary porcine alveolar macrophages (PAMs) revealed that rASFV-Gluc/EGFP exhibits similar biological characteristics to ASFV-WT. Furthermore, analysis of Gluc activities, fluorescence, and next-generation sequencing indicated that rASFV-Gluc/EGFP maintains good genetic stability after 20 consecutive passages in PAMs. Using the HTS platform established with rASFV-Gluc/EGFP, we screened and identified phenazine-1-carboxylic acid (PCA) as an effective inhibitor of ASFV replication from 246 small molecule compounds in PAMs. Importantly, PCA was found to reduce ASFV replication by as much as 100-fold at a concentration of 25 μM. Overall, this study suggests that rASFV-Gluc/EGFP is suitable for rapid screening of anti-ASFV drugs. Importantly, we showed that PCA has significant anti-ASFV activity in PAMs.https://doi.org/10.1186/s13567-025-01467-2African swine feverAfrican swine fever virusdual-reporter virushigh-throughput screeningphenazine-1-carboxylic acid |
| spellingShingle | Jing Lan Rui Luo Di Liu Changxing Qi Xin Song Zhanhao Lu Ruojia Huang Yuying Yang Yuan Sun Yonghui Zhang Tao Wang Hua-Ji Qiu A novel high-throughput screen identifies phenazine-1-carboxylic acid as an inhibitor of African swine fever virus replication in primary porcine alveolar macrophages Veterinary Research African swine fever African swine fever virus dual-reporter virus high-throughput screening phenazine-1-carboxylic acid |
| title | A novel high-throughput screen identifies phenazine-1-carboxylic acid as an inhibitor of African swine fever virus replication in primary porcine alveolar macrophages |
| title_full | A novel high-throughput screen identifies phenazine-1-carboxylic acid as an inhibitor of African swine fever virus replication in primary porcine alveolar macrophages |
| title_fullStr | A novel high-throughput screen identifies phenazine-1-carboxylic acid as an inhibitor of African swine fever virus replication in primary porcine alveolar macrophages |
| title_full_unstemmed | A novel high-throughput screen identifies phenazine-1-carboxylic acid as an inhibitor of African swine fever virus replication in primary porcine alveolar macrophages |
| title_short | A novel high-throughput screen identifies phenazine-1-carboxylic acid as an inhibitor of African swine fever virus replication in primary porcine alveolar macrophages |
| title_sort | novel high throughput screen identifies phenazine 1 carboxylic acid as an inhibitor of african swine fever virus replication in primary porcine alveolar macrophages |
| topic | African swine fever African swine fever virus dual-reporter virus high-throughput screening phenazine-1-carboxylic acid |
| url | https://doi.org/10.1186/s13567-025-01467-2 |
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