可作为疫苗佐剂的云芝多糖磷酸钙纳米粒的制备与表征  

Preparation and Characterization of Polysaccharide Krestin Calcium Phosphate(PSK-CAP) Nanoparticle as a Vaccine Adjuvant

作　　者：宋志磊[1] 潘玉涛[1] 黄新河[1] 刘新荣[1] 晏为力[1] SONG Zhi-lei PAN Yu-tao HUANG Xin-he LIU Xin-rong YAN Wei-li(School of life science and engineering, Southwest Jiaotong University, Chengdu 610031, China)

机构地区：[1]西南交通大学生命科学与工程学院,成都610031

出　　处：《天然产物研究与开发》2016年第9期1357-1362,共6页Natural Product Research and Development

基　　金：四川省科技厅项目(2015JY0202);成都市科技惠民项目(2015-HM01-00047-SF)

摘　　要：我们设想通过磷酸钙药物载体与多糖的结合制备新型的疫苗佐剂,以提高疫苗传递效率,增强免疫应答。首先对云芝粗多糖进行提纯处理,随后利用高碘酸钠(Na IO4)氧化云芝多糖(PSK)得到多醛基产物,再以帕米膦酸二钠(PA)修饰,得到PSK-PA,利用红外光谱法对其产物进行鉴定。采用共沉淀法,制备功能化磷酸钙(PSK-CAP)纳米粒,利用动态光散射(DLS)粒度仪对纳米粒性能进行对比分析选取最优结果,并通过扫描电子显微镜(SEM)观察最优条件下纳米粒的形貌。随后利用载有鸡卵清蛋白(OVA)为模型抗原的纳米粒(PSKCAP-OVA)验证对细胞活性影响,同时观察RAW264.7细胞对其吞噬能力。通过一系列的表征发现,纳米粒平均粒径为100 nm左右,且分散性良好,PSK-CAP-OVA纳米粒毒性较小,并且RAW264.7细胞更倾向于吞噬装载OVA的纳米粒,可以提高胞内OVA的浓度。总之,PSK-CAP纳米粒可为后续研究佐剂类型提供参考。We hypothesize that calcium phosphate（ CAP） coated polysaccharide as vaccine adjuvant can improve the transmission efficiency as well as enhance immune response. In this study,unpurified polysaccaride krestins（ PSK） was treated in traditional craft. PSK was then oxidized by sodium periodate to obtain a multi-aldehyde product. PSK-aldehyde was reacted with pamidronate（ PA） to obtain Pa-modified PSK（ PSK-PA） and the products were identified by infrared spectrum（ IR）. PSK-PA was used to synthesize CAP nanoparticles by co-precipitation method. The size and PDI was used to optimize the formulation of the nanoparticle. The morphology of nanoparticles was observed using scanning electron microscope（ SEM）. Subsequently,ovalbumin（ OVA）,used as a model antigen,was loaded into the nanoparticles to examine the cytotoxicity on RAW264. 7. The uptake of PSK-CAP-OVA-FITC loaded nanoparticle was investigated on RAW264. 7 by fluorescent microscopy. The average diameter of the nanoparticles was about 100 nm with a PDI less than0. 3 indicating a homogeneous dispersion. Interestingly,the toxicity of PSK-CAP-OVA nanoparticles was significantly reduced when compared to the free PA. Additionally,we observed that Raw264. 7 cell preferentially uptake the PSK-CAPOVA nanoparticles compared to that of the free OVA. In conclusion,PSK-CAP nanoparticles can provide a reference for future studies on immune adjuvants.

关 键 词：云芝多糖 磷酸钙纳米粒 免疫佐剂 OVA 纳米粒表征 

分 类 号：R944.1[医药卫生—药剂学]