宿主因子NPSR调节伪狂犬病病毒感染能力的研究OA北大核心CSTPCD
Regulation of pseudorabies virus infectivity by the host factor NPSR
[目的]伪狂犬病病毒(Pseudorabies virus,PRV)属甲型疱疹病毒科,是一种嗜神经性病毒,通过接触传播感染造成神经和呼吸系统疾病.猪是PRV的自然宿主,PRV的感染和流行对养猪业造成巨大经济损失.PRV侵染宿主中枢神经系统后,引发神经肽S(Neuropeptide S,NPS)系统表达水平的改变.NPS是一种神经肽物质,作为神经内分泌系统的关键成员在中枢神经系统中广泛分布,与慢性疾病相关联,NPS通过神经肽S受体(Neuropeptide S receptor,NPSR)介导多样化的生理功能,在机体内以NPS/NPSR系统形式共同参与神经内分泌-免疫网络调节.本研究探讨了NPS/NPSR系统在PRV感染宿主细胞中的作用,以期待开发新的抗PRV靶点.[方法]通过CRISPR/Cas9 构建NPS和NPSR敲除的PK15 细胞系,利用qPCR、免疫荧光、Western blot、免疫共沉淀(CO-IP)等方法检测感染PRV病毒后细胞系的抗病毒能力及NPSR与病毒表面糖蛋白的结合能力.[结果]NPSR敲除能减弱PRV对PK15 细胞的感染能力,而NPS基因敲除增强了PRV对PK15 细胞的感染能力.NPSR敲除细胞在PRV吸附过程中能减少病毒的侵入量,因此NPSR是潜在的PRV入侵受体.CO-IP试验证明,NPSR与病毒包膜蛋白gB和gD存在相互作用.[结论]NPSR通过与病毒表面糖蛋白结合促进PRV感染,NPSR可能是PRV感染神经系统的受体因子之一.NPSR敲除能显著抑制PRV对PK15 细胞的感染.NPSR可为开发PRV防控药物和治疗策略提供新的研究靶点.
[Objective]Pseudorabies virus(PRV),belonging to the herpesviridae family,is a neurotropic virus that spreads infection and causes neurological and respiratory diseases through contact.Pigs are the natural hosts of PRV,and PRV infections and epidemics cause great economic losses to the pig industry.PRV infection of the host central nervous system triggers changes in the expression level of the neuropeptide S(NPS)system,a neuropeptide substance that is widely distributed in the central nervous system as a key member of the neuroendocrine system,and is associated with chronic diseases.NPS mediates diverse physiological functions through the neuropeptide S receptor(NPSR),which participates in the regulation of the neuroendocrine-immune network in the body as the NPS/NPSR system together.In this study,we investigated the role of the NPS/NPSR system in PRV-infected host cells in anticipation of developing new anti-PRV targets.[Method]NPS and NPSR knock-out PK15 cell lines were constructed by CRISPR/Cas9 method.The antiviral ability of cell lines infected with PRV virus and the binding ability of NPSR to viral surface glycoproteins were detected by qPCR,immunofluorescence,Western blot and co-immunoprecipitation(CO-IP).[Result]NPSR knockout attenuated the infectivity of PRV on PK15 cells,while NPS knockout enhanced the infectivity of PRV on PK15 cells.NPSR knockout cells reduced viral invasion during PRV adsorption,thus NPSR was a potential PRV invasion receptor.The interaction of NPSR with viral envelope proteins gB and gD were demonstrated by CO-IP experiment.[Conclusion]NPSR promotes PRV infection by binding to viral surface glycoproteins,and NPSR may be one of the receptor factors for PRV infection of the nervous system.NPSR knockout significantly inhibits the infection of PK15 cells by PRV.NPSR may provide new research targets for the development of drugs and therapeutic strategies for PRV prevention and control.
罗欣欣;郑虎;高美娟;杨化强
华南农业大学动物科学学院/国家生猪种业工程技术中心,广东广州 510642
畜牧业
神经肽S受体伪狂犬病病毒病毒受体宿主病毒相互作用
Neuropeptide S receptorPseudorabies virusVirus receptorHost-virus interaction
《华南农业大学学报》 2024 (004)
457-468 / 12
国家自然科学基金(32372874)
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