Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, 360 Huntington Avenue, Boston, MA 02115, USA.
Int J Pharm. 2013 Sep 10;453(2):400-7. doi: 10.1016/j.ijpharm.2013.05.028. Epub 2013 May 21.
The purpose of this study was to develop a water-in-oil-in-water (W/O/W) multiple emulsions-based vaccine delivery system for plasmid DNA encoding the gp100 peptide antigen for melanoma immunotherapy. The gp100 encoding plasmid DNA was encapsulated in the inner-most aqueous phase of squalane oil containing W/O/W multiple emulsions using a two-step emulsification method. In vitro transfection ability of the encapsulated plasmid DNA was investigated in murine dendritic cells by transgene expression analysis using fluorescence microscopy and ELISA methods. Prophylactic immunization using the W/O/W multiple emulsions encapsulated the gp100 encoding plasmid DNA vaccine significantly reduced tumor volume in C57BL/6 mice during subsequent B16-F10 tumor challenge. In addition, serum Th1 cytokine levels and immuno-histochemistry of excised tumor tissues indicated activation of cytotoxic T-lymphocytes mediated anti-tumor immunity causing tumor growth suppression. The W/O/W multiple emulsions-based vaccine delivery system efficiently delivers the gp100 plasmid DNA to induce cell-mediated anti-tumor immunity.
本研究旨在开发一种基于水包油包水（W/O/W）多重乳液的疫苗传递系统，用于递送编码黑色素瘤免疫治疗用 gp100 肽抗原的质粒 DNA。使用两步乳化法，将编码 gp100 的质粒 DNA 包封在含有 W/O/W 多重乳液的 squalane 油的最内侧水相中。通过荧光显微镜和 ELISA 方法进行转染表达分析，研究了包封的质粒 DNA 在小鼠树突状细胞中的体外转染能力。使用 W/O/W 多重乳液包封 gp100 编码质粒 DNA 疫苗进行预防性免疫接种，可显著减少随后 B16-F10 肿瘤攻击时 C57BL/6 小鼠的肿瘤体积。此外，血清 Th1 细胞因子水平和切除的肿瘤组织免疫组织化学分析表明，细胞毒性 T 淋巴细胞介导的抗肿瘤免疫被激活，导致肿瘤生长抑制。基于 W/O/W 多重乳液的疫苗传递系统可有效传递 gp100 质粒 DNA，从而诱导细胞介导的抗肿瘤免疫。
