作为疫苗佐剂载体的β-葡聚糖颗粒
Beta-Glucan Particles as Vaccine Adjuvant Carriers.
作者信息
Mirza Zeynep, Soto Ernesto R, Dikengil Fusun, Levitz Stuart M, Ostroff Gary R
机构信息
Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
Department of Medicine, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
出版信息
Methods Mol Biol. 2017;1625:143-157. doi: 10.1007/978-1-4939-7104-6_11.
Glucan particles (GPs) are spherical hollow particles derived from Saccharomyces cerevisiae cell walls and mainly consist of β-1, 3-D-glucans. The inner hollow cavity of glucan particles can be loaded with different compounds, including protein antigens, and delivered to macrophages and dendritic cells. Moreover, the GP delivery system possesses β-glucan's intrinsic immunostimulatory properties. Therefore, GPs serve as both an antigen-presenting cell-targeted delivery system and an adjuvant.Here, we describe the production of GPs from S. cerevisiae using hot alkaline and solvent extraction and characterization of these particles for morphology, particle density, and hydrodynamic volume. A detailed protocol for loading and entrapping a model antigen, ovalbumin (OVA), into these particles using yeast RNA is presented. Similar methods are used to load pathogen-specific antigens (peptides, proteins, soluble extracts) which then can be tested in in vivo vaccination models.
葡聚糖颗粒（GPs）是源自酿酒酵母细胞壁的球形中空颗粒，主要由β-1,3-D-葡聚糖组成。葡聚糖颗粒的内部中空腔可装载不同的化合物，包括蛋白质抗原，并递送至巨噬细胞和树突状细胞。此外，GP递送系统具有β-葡聚糖固有的免疫刺激特性。因此，GPs既作为靶向抗原呈递细胞的递送系统，又作为佐剂。在此，我们描述了使用热碱性和溶剂萃取法从酿酒酵母中生产GPs，并对这些颗粒的形态、颗粒密度和流体动力学体积进行表征。还介绍了使用酵母RNA将模型抗原卵清蛋白（OVA）装载并包封到这些颗粒中的详细方案。采用类似方法装载病原体特异性抗原（肽、蛋白质、可溶性提取物），然后可在体内疫苗接种模型中进行测试。
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