枸杞多糖超大介孔二氧化硅的黏膜免疫佐剂活性及其作用机制研究

新型黏膜免疫佐剂的开发对于动物疫病的防控具有重要意义。前期研究发现脂质体存在渗漏、在胃肠道中不稳定等缺陷。而超大介孔二氧化硅（UCMS）更灵活、坚固、耐pH，具有大的孔体积和良好的生物相容性等优良性质，可以保护抗原不被胃肠道消化液降解。因此UCMS作为枸杞多糖（LBP）的递送载体对于新型黏膜免疫佐剂的开发更具优势。本项目通过优化制备得到质量稳定、抗原负载率高的LBP-UCMS；然后将抗原（H9N2亚型禽流感灭活病毒）负载于LBP-UCMS，口服免疫雏鸡，检测机体黏膜免疫应答和全身免疫应答水平；采用流式细胞仪、激光共聚焦、蛋白质组学、western blotting等技术，探究LBP-UCMS靶向递送抗原并通过CX3CR1、TLR2、TLR4、NOD1、NOD2受体途径激活肠黏膜派尔集合淋巴结树突状细胞，阐明其发挥黏膜佐剂活性的机制。将为中药多糖新型黏膜佐剂的研制及动物疫病的的防控提供参考。

The development of new mucosal immune adjuvants is of great significance for the prevention and control of animal diseases. Previous studies have found that liposomes have leakage, instability in the gastrointestinal tract and other defects. However, the ultra-large pore mesoporous silica (UCMS) is more flexible, strong, pH resistant, with large pore volume and good biocompatibility and other excellent properties, which can protect the antigen from degradation by gastrointestinal digestive fluid. Therefore, UCMS as delivery carrier of Lycium barbarum polysaccharides (LBP) has more advantages for the development of new mucosal immune adjuvants. In this project, LBP-UCMS with stable quality and high antigen loading rate will be obtained through optimized preparation. Then the antigen (H9N2 subtype avian influenza inactivated virus) will be loaded in LBP-UCMS, and chicks will be immunized orally to detect the level of mucosal immune response and systemic immune response. Using flow cytometry, laser confocal, proteomics, western blotting and other technologies to explore the mechanism of LBP-UCMS targeting antigen delivery and activation of intestinal Peyer’s patch lymph node dendritic cells through CX3CR1, TLR2, TLR4, NOD1, NOD2 receptor pathway, elucidating its mechanism of mucosal adjuvant activity. It will provide a reference for the research and development of new mucosal adjuvant of Chinese medicine polysaccharide and the prevention and control of animal disease.