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口服重组酿酒酵母向小鼠树突状细胞内的基因传递。

Gene delivery to dendritic cells by orally administered recombinant Saccharomyces cerevisiae in mice.

机构信息

College of Animal Science & Technology, Shaan'xi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, YangLing, Shaan'xi 712100, PR China.

出版信息

Vaccine. 2013 Feb 27;31(10):1360-3. doi: 10.1016/j.vaccine.2012.11.048. Epub 2012 Nov 28.

Abstract

DNA vaccination has caught the attention of many for triggering humoral as well as cellular immune responses. And delivering DNA into the antigen presenting cells (APCs) in order to induce efficient immunoresponse has become the backbone of this field. It has been confirmed that Saccharomyces cerevisiae, though non-pathogenic, is being engulfed by the dendritic cells and macrophages and delivers not only proteins, but also DNA materials (already confirmed in vitro). In this research, S. cerevisiae is used to deliver green fluorescent protein (GFP) reporter gene controlled under cytomegalovirus (CMV) promoter in living organism (mice). The recombinant yeast, transfected with the plasmid containing the GFP gene, was heat killed and orally administered to mice. After 60 h of yeast administration, mice were sacrificed and intestine was separated, washed and frozen in liquid nitrogen. Tissues were cut at the size of 10 μm using Cryostat machine, and GFP expression was successfully detected under a fluorescence microscope. After 45 days Western blot was able to detect GFP antibody in the blood of mice. These results imply that S. cerevisiae, being non-pathogenic, cheap, and easy to culture could be a good candidate to deliver DNA materials to the immune cells for vaccination.

摘要

DNA 疫苗引起了许多人的关注,因为它可以引发体液和细胞免疫反应。为了诱导有效的免疫反应,将 DNA 递送到抗原呈递细胞(APCs)中已成为该领域的核心。已经证实,酵母虽然是非致病性的,但被树突状细胞和巨噬细胞吞噬,并不仅传递蛋白质,还传递 DNA 物质(已在体外得到证实)。在这项研究中,酵母被用于在活生物体(小鼠)中递送受巨细胞病毒(CMV)启动子控制的绿色荧光蛋白(GFP)报告基因。用含有 GFP 基因的质粒转染的重组酵母经热处理后口服给予小鼠。酵母给药 60 小时后,处死小鼠并分离、清洗和在液氮中冷冻肠道。使用冷冻切片机制备大小为 10 μm 的组织切片,并在荧光显微镜下成功检测到 GFP 表达。45 天后,Western blot 能够在小鼠血液中检测到 GFP 抗体。这些结果表明,酵母作为一种非致病性、廉价且易于培养的候选物,可以将 DNA 物质递送到免疫细胞中用于疫苗接种。

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