香菇多糖氧化石墨烯佐剂活性及其对巨噬细胞作用分子机理的研究

香菇多糖作为免疫增强剂可以增强动物的免疫力，但是其存在体内代谢快、靶向性差等缺点，一定程度上限制了其作为疫苗佐剂的应用。因此，为了提高香菇多糖的利用率和靶向性，本研究拟将香菇多糖负载在氧化石墨烯的表面实现香菇多糖和抗原的共同装载和协同递送，增强香菇多糖的靶向性，提高香菇多糖的免疫增强作用。首先，通过优化制备得到质量稳定的香菇多糖氧化石墨烯。然后，将香菇多糖氧化石墨烯作为猪圆环病毒疫苗的佐剂免疫仔猪，比较免疫后仔猪的体液免疫和细胞免疫反应水平，并观察注射部位与淋巴结中免疫细胞募集、激活情况，分析局部免疫与整体免疫水平的相关性，阐明其体内佐剂活性作用机理。最后，探究香菇多糖氧化石墨烯通过Integrin、TLR2、TLR4或Dectin-1途径活化巨噬细胞发挥佐剂效应的作用机制，并通过基因敲除动物模型进行体内验证，最终全面阐明香菇多糖氧化石墨烯发挥佐剂作用的机制，为其临床应用提供依据。

Lentinan, as an immunopotentiator, coucld enhance the immunity of animals. However, some shortcomings, such as rapid metabolism and poor targeting in vivo, have limited its application as vaccine adjuvant. Therefore, in this study, Lentinan will be loaded on the surface of graphene oxide to achieve co-loading and synergistic delivery of Lentinan and antigen, which can enhance the targeting and immune enhancement of Lentinan. Firstly, stable Lentinan graphene oxide will be prepared by optimization. Secondly, the levels of humoral and cellular immune response of lentinan graphene oxide on porcine circovirus vaccines will be compared as adjuvant. The association between local immunity and overall immunity will be analyzed by detecting the recruitment and activation of immune cells in the injection site and lymph node after immunization to elucidate the mechanism of adjuvant activity in vivo. Finally, the effect of lentinan graphene oxide on macrophages will be analyzed to elucidate the mechanism of lentinan graphene oxide on the activation of macrophage by Integrin, TLR2, TLR4 or Dectin-1 pathway. And the results will be verified by gene knockout animal models. The adjuvant activity mechanism of lentinan graphene oxide will be explained, which will provide a basis for further clinical application.