锰纳米颗粒的制备及其增强免疫功能研究

Manganese nanoparticles: preparation and application as an immunopotentiator

摘要

[目的]本文旨在研究锰纳米颗粒(Mn^nan)的制备方法，进一步探究其对抗原递呈细胞的激活及对 H9N2 疫苗的免疫增强效力。[方法]首先用鸡血清白蛋白和氯化锰制备了锰纳米颗粒，然后通过纳米颗粒相关表征方法对锰纳米颗粒进行了鉴定，最后将其作为免疫增强剂，分析其对抗原递呈细胞的激活和对 H9N2 疫苗的免疫增强效果。[结果]利用鸡血清白蛋白和氯化锰制备的锰纳米颗粒粒径为 50nm 左右，呈立方体状，4℃放置 60d 仍呈现较好的稳定性。X 射线衍射(XRD)分析显示制备的锰纳米颗粒为四氧化三锰(Mn₃O₄)。体外试验显示，锰纳米颗粒能够激活 RAW-Blue 细胞的 NF-κB 信号通路，并且诱导了 IFN-γ、IFN-β、IL-12、TNF-α mRNA 的表达。通过免疫鸡后发现锰纳米颗粒能提高 H9N2 病毒免疫后特异性的血凝抗体水平。[结论]成功制备了锰纳米颗粒，证实了其在体内外均有良好的免疫刺激活性。研究结果为兽用纳米级免疫增强剂的研发奠定了基础。

Abstract

[Objectives] The paper aimed to study the activation of manganese nanoparticles (Mn^nan) on antigen presenting cells and the immune enhancing efficacy of the H9N2 vaccine. [Methods] This research prepared nanoparticles with chicken serum albumin and manganese chloride, and then identified manganese nanoparticles by nanoparticle correlation characterization method. Finally, manganese nanoparticles were used as immunopotentiator. The activation of antigen presenting cells and the immune enhancing efficacy of the H9N2 vaccine were analyzed. [Results] The size of manganese nanoparticles was about 50 nm, and the shape was like a cube. It showed good stability after being placed at 4 ℃ for 60 days. X-ray diffraction (XRD) data showed that the prepared manganese nanoparticles were manganic tetroxide. In vitro experiments showed that manganese nanoparticles could activate the NF-κB signaling pathway of RAW-Blue cells and induce the expression of IFN-γ, IFN-β, IL-12 and TNF-α mRNA. Manganese nanoparticles could increase the specific hemagglutination inhibition antibody level after H9N2 virus immunization in chickens. [Conclusions] Manganese nanoparticles were successfully prepared in this study, which proved that they had good immunostimulating activity in vivo and in vitro. The results of this study laid a foundation for the research and development of nanoscale immunopotentiator for veterinary use.

关键词

锰纳米颗粒 / 免疫增强剂 / 抗原递呈细胞的激活 / H9N2 灭活苗

Key words

manganese nanoparticle / immunopotentiator / activation of antigen-presenting cells / H9N2 inactivated vaccine

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基金资助

国家重点研发计划项目(2022YFD1800800)

江苏省农业自主创新专项(CX(24)3079)