受体介导的金纳米复合物靶向阻断及光热杀灭猪伪狂犬病病毒研究

Pseudorabies is an acute and severe infectious disease caused by porcine pseudorabies virus (PRV) infection, which is a neurotropic and latently infectious pathogen. And its near 100% mortality rate has always been a major problem in virology and aquaculture. In our project, PRV strain QHD-1 was used as a model, and the receptor-mediated gold nanocomposites (Nectin-1/AuNPs) were prepared based on the Escherichia coli prokaryotic expression system and chemical modification to specifically recognize and neutralize the viruses, thus blocking the invasion of viruses into cells. The recognition effect can further induce the gold nanoparticles to arrange orderly on the surface of the virus, so as to construct the self-assembly of gold nanoparticles centering on the virus and realize the photothermal killing of the virus. Laser confocal fluorescence microscopy, thermal imager and high resolution fluorescence microscopy were used to evaluate the inhibition efficiency on viral titer and invasion process in vitro and in vivo, and to elucidate the mechanism of targeted photothermal killing of viruses mediated by cell receptors. This project provides a new strategy for pre-infection and post-infection targeted treatment of pseudorabies virus, and has important reference significance for other virus control studies.

猪伪狂犬病是由猪伪狂犬病病毒（PRV）感染引起的嗜神经性和潜伏感染严重的急性、烈性传染病，其近乎100%的致死率一直是困扰病毒学和养殖业的重大难题。本项目拟以PRV强毒株QHD-1为研究毒株，基于大肠杆菌原核表达系统和化学修饰等手段，制备受体介导的金纳米复合物（Nectin-1/AuNPs）用于特异性识别与中和病毒，从而阻断病毒入侵细胞。该识别作用可进一步诱导金纳米粒子在病毒表面有序排列，从而构建以病毒为中心的金纳米粒子自组装体，实现对病毒的光热杀灭。采用激光共聚焦荧光显微镜、热成像仪和高分辨荧光显微成像等技术，从细胞和活体水平评价其对病毒滴度、侵入过程的抑制效率，阐明细胞受体介导的靶向光热杀灭病毒机制。本项目为猪伪狂犬病的感染前阻断和感染后定向治疗提供了新的策略，对其他病毒的防治研究也具有重要的参考意义。

病原微生物对人类健康和全球公共卫生安全存在潜在的安全隐患，寻找新型有效的防治措施具有重大科学意义与应用价值，而纳米药物的出现为病原微生物的精准防控带来了新的希望。猪伪狂犬病是由猪伪狂犬病病毒（PRV）感染引起的嗜神经性和潜伏感染严重的急性、烈性传染病，其近乎100%的致死率一直是困扰病毒学和养殖业的重大难题。紧急免疫接种和控制继发感染是目前防治伪狂犬病的主要策略，然而一旦动物出现临床发病症状时，尚无确切有效的针对性治疗措施。因此，探索制备靶向、高效杀灭伪狂犬病病毒的新材料和新方法迫在眉睫。针对上述问题，本项目基于受体与病毒糖蛋白间的特异性相互作用以及纳米策略，从制备高效控制病原微生物感染的纳米药物以及探究机制方面开展相关研究工作，具体如下：（1）通过大肠杆菌原核表达系统成功表达了具有活性的PRV受体蛋白Nectin-1，可有效结合PRV并抑制PRV的感染；（2）制备了具有光热活性的GO@AuNPs复合材料，实现了对猪伪狂犬病病毒的感染前阻断和感染后定性治疗；（3）制备了金银混合纳米棒，通过调控病毒感染过程中ROS的产生来定量释放银离子，实现了长效的抗病毒作用；（4）构建了光响应性纳米载药系统、纳米复合水凝胶以及非抗生素纳米复合物，实现了对病原微生物的高效清除。本项目为病原微生物的精准治疗提供了新的策略。在课题执行过程中，项目负责人以一作、并列一作或并列通讯发表SCI论文6篇，申请专利4项，已授权1项，参与相关学术会议3次。