Insect-specific neurotoxic peptides derived from the venoms of scorpions and spiders can cause acute paralysis and death when injected into insects,offering a promising insecticidal component for insect pest control.H...Insect-specific neurotoxic peptides derived from the venoms of scorpions and spiders can cause acute paralysis and death when injected into insects,offering a promising insecticidal component for insect pest control.However,effective delivery systems are required to help neurotoxic peptides pass through the gut barrier into the hemolymph,where they can act.Here,we investigated the potential of a novel nanocarrier,Drosophila X virus-like particle(DXV-VLP),for delivering a neurotoxin from the scorpion Androctonus australis Hector(AaIT)against the invasive pest fruit fly,Drosophila suzuki.Our results show that the fusion proteins of DXV polyproteins with AalT peptide at their Ctermini could be sufficiently produced in Lepidoptera Hi5 cells in a soluble form using the recombinant baculovirus expression system,and could self-assemble into VLPs with similar particle morphology and size to authentic DXV virions.In addition,the AalT peptides displayed on DXV-VLPs retained their toxicity,as demonstrated in injection bioassays that resulted in severe mortality(72%)in adults after 72 h.When fed to adults,mild mortality was observed in the group treated with DXV-AalT(38%),while no mortality occurred in the group treated with AaIT peptide,thus indicating the significant role of DXV-VLPs in delivering AalT peptides.Overall,this proof-of-concept study demonstrates for the first time that VLPs can be exploited to enhance oral delivery of insect-specific neurotoxic peptides in the context of pest control.Moreover,it provides insights for further improvements and potentially the development of neurotoxin-based bioinsecticides and/or transgenic crops for insect pest control.展开更多
Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes high fatality.The nucleocapsid of CCHFV is comprised of N prote...Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes high fatality.The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment.In this research,the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus.Under an electron microscope,Virus-Like Particles (VLPs) with various size and morphology were observed in cytoplasmic vesicles in the infected cells.Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation,which was confirmed by Western blot analysis and immuno-electron microscopy (IEM).展开更多
病毒样颗粒(virus like particles,VLPs)近年来被广泛应用于疫苗和治疗性药物递送系统领域。与体内组装VLPs相比,体外组装VLPs具有组装寡聚体成分明确、反应条件可控、避免宿主成分的引入以及能实现药物活性成分的包裹和递送功能等诸多...病毒样颗粒(virus like particles,VLPs)近年来被广泛应用于疫苗和治疗性药物递送系统领域。与体内组装VLPs相比,体外组装VLPs具有组装寡聚体成分明确、反应条件可控、避免宿主成分的引入以及能实现药物活性成分的包裹和递送功能等诸多优势。概述了体外VLPs的生产方法、影响体外VLPs形成的因素、VLPs的特征分析方法,综述了体外VLPs在疫苗和治疗性药物递送方面的应用进展,期望对体外VLPs组装技术的发展提供参考思路,促进体外VLPs组装技术在疾病预防和治疗应用方面发挥更大的作用。展开更多
基金supported by the Special Research Fund of Ghent University and Research Foundation Flanders(FWO).CNTT is recipient of a senior postdoctoral fellowship from FWO(grant number 12V5722N).
文摘Insect-specific neurotoxic peptides derived from the venoms of scorpions and spiders can cause acute paralysis and death when injected into insects,offering a promising insecticidal component for insect pest control.However,effective delivery systems are required to help neurotoxic peptides pass through the gut barrier into the hemolymph,where they can act.Here,we investigated the potential of a novel nanocarrier,Drosophila X virus-like particle(DXV-VLP),for delivering a neurotoxin from the scorpion Androctonus australis Hector(AaIT)against the invasive pest fruit fly,Drosophila suzuki.Our results show that the fusion proteins of DXV polyproteins with AalT peptide at their Ctermini could be sufficiently produced in Lepidoptera Hi5 cells in a soluble form using the recombinant baculovirus expression system,and could self-assemble into VLPs with similar particle morphology and size to authentic DXV virions.In addition,the AalT peptides displayed on DXV-VLPs retained their toxicity,as demonstrated in injection bioassays that resulted in severe mortality(72%)in adults after 72 h.When fed to adults,mild mortality was observed in the group treated with DXV-AalT(38%),while no mortality occurred in the group treated with AaIT peptide,thus indicating the significant role of DXV-VLPs in delivering AalT peptides.Overall,this proof-of-concept study demonstrates for the first time that VLPs can be exploited to enhance oral delivery of insect-specific neurotoxic peptides in the context of pest control.Moreover,it provides insights for further improvements and potentially the development of neurotoxin-based bioinsecticides and/or transgenic crops for insect pest control.
基金The Knowledge Innovation Program of the Chinese Academy of Sciences,Grant No.KSCX2-YW-N-065the Knowledge Innovation Program of the Chinese Academy of Sciences,Grant No.KSCX2-EW-G-8+1 种基金National Key Basic Research Program(973Program),Grant No.2010CB530103National Basic Research Priorities Program of China,Grant No.2007FY210700
文摘Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes high fatality.The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment.In this research,the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus.Under an electron microscope,Virus-Like Particles (VLPs) with various size and morphology were observed in cytoplasmic vesicles in the infected cells.Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation,which was confirmed by Western blot analysis and immuno-electron microscopy (IEM).
文摘病毒样颗粒(virus like particles,VLPs)近年来被广泛应用于疫苗和治疗性药物递送系统领域。与体内组装VLPs相比,体外组装VLPs具有组装寡聚体成分明确、反应条件可控、避免宿主成分的引入以及能实现药物活性成分的包裹和递送功能等诸多优势。概述了体外VLPs的生产方法、影响体外VLPs形成的因素、VLPs的特征分析方法,综述了体外VLPs在疫苗和治疗性药物递送方面的应用进展,期望对体外VLPs组装技术的发展提供参考思路,促进体外VLPs组装技术在疾病预防和治疗应用方面发挥更大的作用。
