The rapid expansion of large-scale pig farming has brought about a surge in viral diseases with high morbidity rates and diverse manifestations.This widespread occurrence of multiple viral diseases in pig farms has in...The rapid expansion of large-scale pig farming has brought about a surge in viral diseases with high morbidity rates and diverse manifestations.This widespread occurrence of multiple viral diseases in pig farms has inflicted severe economic losses on the global swine industry.Consequently,there is an urgent need for eco-friendly and efficient antivi-ral drugs that can effectively combat viruses and prevent diseases such as PEDV,PRRSV,PRV,and other viral infections.To this end,we conducted a study on the antiviral activity and cytotoxicity of eleven different Chinese herbal extracts(CHE) against PRV.In vitro testing of several extracts,namely,Echinacea,Ilex purpurea Hassk,Ganoderma lucidum Kars,Taraxacum mongolicum,and Ilex rotunda Thunb,exhibited remarkable inhibition of PRV infection without causing any cytotoxic effects.Specifically,their antiviral selectivity indexes were significantly higher,with values ranging from 6-to 144-fold.The antiviral efficacy of five CHEs was evaluated against other RNA viruses,including PRRSV and PEDV.The extracts showed substantial inhibition of PEDV and PRRSV proliferation.Echinacea and Ilex purpurea Hassk extracts exhibited the highest virus inhibitory effects.To understand the antiviral mechanisms underlying their potent activity,a time-of-addition experiment was conducted.The results indicated that these extracts effectively targeted the early infection and postinfection stages of PRV,PEDV,and PRRSV.The study found that the Chinese herbal extracts,Echinacea and Ilex purpurea Hassk,had both direct and indirect effects on virus particles and cellular targets,demonstrating broad-spectrum antiviral activity against multiple clinical strains of PRV and PEDV.These findings provide a strong foundation for the development of herbal medicines to prevent and treat infections caused by PRV,PEDV and PRRSV in the swine industry.The identified extracts show great promise for the formulation of effective and environmentally friendly antiviral interventions.展开更多
Porcine epidemic diarrhea virus(PEDV),an enteric coronavirus,is widely spread worldwide and causes huge economic losses.The effective measure to control the viral infection is to develop ideal vaccines.Here,the collag...Porcine epidemic diarrhea virus(PEDV),an enteric coronavirus,is widely spread worldwide and causes huge economic losses.The effective measure to control the viral infection is to develop ideal vaccines.Here,the collagenase equivalent domain(COE)of PEDV was displayed on the surface of nanoparticles(NPs)in order to develop a newer,safer and more effective subunit vaccine against PEDV.The monomeric COE was displayed on the mi3 protein,which self-assembles into nanoparticles composed of 60 subunits,using the SpyTag/SpyCatcher system.The size,zeta potential,microstructure of the COE-mi3 virus-like particles(VLPs)were investigated.The COE-mi3 VLPs that possessed good security,stability and better retention can be more efficiently taken up by antigen-presenting cells(APCs)and help promote dendritic cells(DCs)maturation.Moreover,COE-mi3 VLPs could prominently improve specifc antibody levels including neutralizing antibodies(NAbs),and serum IgG,mucosal IgA.Moreover,COE-mi3 VLPs elicited more activation of CD4^(+)and CD8^(+)T cells and production of IFN-γand IL-4 cytokines.In particular,COE-mi3 VLPs is an effectual antigen-delivery platform to enhance germinal center(GC)B cell responses.This structure-based self-assembly of NP gives great potential to be developed as a new subunit vaccines attractive platform,and may also provide new ideas for the development of other enteric coronavirus vaccines.展开更多
Porcine epidemic diarrhea (PED), caused by porcine epidemic diarrhea virus (PEDV), is a highly contagious gastrointestinal disease characterized by vomiting, diarrhea, and dehydration, with mortality rates approaching...Porcine epidemic diarrhea (PED), caused by porcine epidemic diarrhea virus (PEDV), is a highly contagious gastrointestinal disease characterized by vomiting, diarrhea, and dehydration, with mortality rates approaching 100% among suckling piglets. The PEDV 3C-like protease (3CLpro) is essential for viral replication and regarded as a critical target for antiviral inhibitor development. In this study, we aimed to identify small-molecule inhibitors of PEDV by targeting 3CLpro. Virtual screening of 1.6 million compounds from the ChemDiv library identified four potential candidates. Molecular dynamics simulations, specifically analyzing RMSD, RMSF, and Rg, demonstrated increased structural stability of the compound-protease complexes compared to the monomeric enzyme. All compounds had low cytotoxicity in Vero cells (CC_(50) > 200 μM). Fluorescence resonance energy transfer-based assays demonstrated dose-dependent inhibitory activity of the compounds against 3CLpro. Among the candidates, compound F366-0161 exhibited the weakest inhibition, with an IC_(50) value of 151.5 μM. Two analogues, 3238-0395 (IC_(50) of 121.4 μM) and L878-0493 (IC_(50) of 123.6 μM), exhibited moderately enhanced activity. Y041-1672 was identified as the most effective inhibitor, with an IC_(50) of 86.48 μM. In viral replication inhibition assays, Y041-1672 reduced PEDV replication, with an EC_(50) of 17.97 μM and a selectivity index (SI) of 15.5 (CC_(50) /EC_(50) ). These results were validated by RT-qPCR, plaque assays, immunofluorescence, and Western blot analyses. In vitro validation confirmed Y041-1672 as the optimal antiviral candidate, and time-of-addition experiments indicated that inhibition primarily occurred during viral replication. This study identifies scaffold molecules for PEDV antiviral drug development, providing strategic insights for PED treatment.展开更多
基金supported by the Hubei Agricultural Research System (HBHZDZB-2020-005)the National Natural Science Foundation of China (31872328,32272990)+2 种基金the National Key Research and Development Program of China(2021YFD1800101-2)the Science and Technology Project of Guizhou Province([2020]4Y217)the "Yingzi Tech&Huazhong Agricultural University Intelligent Research Institute of Food Health"(No.IRIFH202209)。
文摘The rapid expansion of large-scale pig farming has brought about a surge in viral diseases with high morbidity rates and diverse manifestations.This widespread occurrence of multiple viral diseases in pig farms has inflicted severe economic losses on the global swine industry.Consequently,there is an urgent need for eco-friendly and efficient antivi-ral drugs that can effectively combat viruses and prevent diseases such as PEDV,PRRSV,PRV,and other viral infections.To this end,we conducted a study on the antiviral activity and cytotoxicity of eleven different Chinese herbal extracts(CHE) against PRV.In vitro testing of several extracts,namely,Echinacea,Ilex purpurea Hassk,Ganoderma lucidum Kars,Taraxacum mongolicum,and Ilex rotunda Thunb,exhibited remarkable inhibition of PRV infection without causing any cytotoxic effects.Specifically,their antiviral selectivity indexes were significantly higher,with values ranging from 6-to 144-fold.The antiviral efficacy of five CHEs was evaluated against other RNA viruses,including PRRSV and PEDV.The extracts showed substantial inhibition of PEDV and PRRSV proliferation.Echinacea and Ilex purpurea Hassk extracts exhibited the highest virus inhibitory effects.To understand the antiviral mechanisms underlying their potent activity,a time-of-addition experiment was conducted.The results indicated that these extracts effectively targeted the early infection and postinfection stages of PRV,PEDV,and PRRSV.The study found that the Chinese herbal extracts,Echinacea and Ilex purpurea Hassk,had both direct and indirect effects on virus particles and cellular targets,demonstrating broad-spectrum antiviral activity against multiple clinical strains of PRV and PEDV.These findings provide a strong foundation for the development of herbal medicines to prevent and treat infections caused by PRV,PEDV and PRRSV in the swine industry.The identified extracts show great promise for the formulation of effective and environmentally friendly antiviral interventions.
基金supported by the Major Scientific and Technological Project of the Henan Province,China(221100110600)the Beijing Life Science Academy,China(2024500CA0010)+1 种基金the Major Program of National Natural Science Foundation of China(32192452)the Chinese Postdoctoral Science Foundation(2023M743209)。
文摘Porcine epidemic diarrhea virus(PEDV),an enteric coronavirus,is widely spread worldwide and causes huge economic losses.The effective measure to control the viral infection is to develop ideal vaccines.Here,the collagenase equivalent domain(COE)of PEDV was displayed on the surface of nanoparticles(NPs)in order to develop a newer,safer and more effective subunit vaccine against PEDV.The monomeric COE was displayed on the mi3 protein,which self-assembles into nanoparticles composed of 60 subunits,using the SpyTag/SpyCatcher system.The size,zeta potential,microstructure of the COE-mi3 virus-like particles(VLPs)were investigated.The COE-mi3 VLPs that possessed good security,stability and better retention can be more efficiently taken up by antigen-presenting cells(APCs)and help promote dendritic cells(DCs)maturation.Moreover,COE-mi3 VLPs could prominently improve specifc antibody levels including neutralizing antibodies(NAbs),and serum IgG,mucosal IgA.Moreover,COE-mi3 VLPs elicited more activation of CD4^(+)and CD8^(+)T cells and production of IFN-γand IL-4 cytokines.In particular,COE-mi3 VLPs is an effectual antigen-delivery platform to enhance germinal center(GC)B cell responses.This structure-based self-assembly of NP gives great potential to be developed as a new subunit vaccines attractive platform,and may also provide new ideas for the development of other enteric coronavirus vaccines.
基金supported by the National Key Research and Development Program of China(2021YFD1800303)the National Natural Science Foundation of China(32473044).
文摘Porcine epidemic diarrhea (PED), caused by porcine epidemic diarrhea virus (PEDV), is a highly contagious gastrointestinal disease characterized by vomiting, diarrhea, and dehydration, with mortality rates approaching 100% among suckling piglets. The PEDV 3C-like protease (3CLpro) is essential for viral replication and regarded as a critical target for antiviral inhibitor development. In this study, we aimed to identify small-molecule inhibitors of PEDV by targeting 3CLpro. Virtual screening of 1.6 million compounds from the ChemDiv library identified four potential candidates. Molecular dynamics simulations, specifically analyzing RMSD, RMSF, and Rg, demonstrated increased structural stability of the compound-protease complexes compared to the monomeric enzyme. All compounds had low cytotoxicity in Vero cells (CC_(50) > 200 μM). Fluorescence resonance energy transfer-based assays demonstrated dose-dependent inhibitory activity of the compounds against 3CLpro. Among the candidates, compound F366-0161 exhibited the weakest inhibition, with an IC_(50) value of 151.5 μM. Two analogues, 3238-0395 (IC_(50) of 121.4 μM) and L878-0493 (IC_(50) of 123.6 μM), exhibited moderately enhanced activity. Y041-1672 was identified as the most effective inhibitor, with an IC_(50) of 86.48 μM. In viral replication inhibition assays, Y041-1672 reduced PEDV replication, with an EC_(50) of 17.97 μM and a selectivity index (SI) of 15.5 (CC_(50) /EC_(50) ). These results were validated by RT-qPCR, plaque assays, immunofluorescence, and Western blot analyses. In vitro validation confirmed Y041-1672 as the optimal antiviral candidate, and time-of-addition experiments indicated that inhibition primarily occurred during viral replication. This study identifies scaffold molecules for PEDV antiviral drug development, providing strategic insights for PED treatment.
