生物
固有层
质粒
乳酸乳球菌
分子生物学
免疫染色
基因传递
微生物学
绿色荧光蛋白
体内
转染
DNA
细菌
乳酸
基因
免疫学
免疫组织化学
上皮
生物化学
生物技术
遗传学
作者
Keita Takahashi,Nozomi Orito,Daiki Yanagisawa,Ayumu Yano,Yusuke Mori,Naoki Inoue
出处
期刊:Vaccine
[Elsevier]
日期:2020-04-01
卷期号:38 (17): 3330-3338
被引量:1
标识
DOI:10.1016/j.vaccine.2020.02.084
摘要
Lactic acid bacteria have been studied as a vehicle for the delivery of plasmid DNA to the gastrointestinal tract. However, low levels of gene expression in vivo limit their practical use. Furthermore, it is still unclear how the orally administrated bacteria transfer their harbored plasmid DNA to host intestinal cells. To more easily track the delivery of plasmid DNA for eukaryotic expression in the intestine, we constructed an L. lactis-E. coli shuttle plasmid (pLEC) that allowed significantly elevated expression of the target protein of interest in eukaryotic cells. We first demonstrated its usefulness for delivery from L. lactis to Caco-2 cells in vitro. We then investigated the cellular target for the L. lactis DNA delivery system in vivo. Mice were orally administrated with LL/pLEC:EGFP, an L. lactis strain carrying pLEC for EGFP expression, and immunofluorescent analyses of frozen sections prepared from their small intestines identified a number of EGFP-expressing cells in the lamina propria and some in the sub-epithelial dome of the Peyer’s patches. Flow cytometric analysis revealed that these EGFP-expressing cells were both CD11c- and F4/80-positive but CX3CR1-negative, suggesting that they are eosinophils. Immunostaining of the sections with an antibody against Siglec-F, a marker protein of eosinophils, confirmed the flow cytometric findings. Thus, the target cells of DNA delivery from L. lactis in the intestines are mainly eosinophils in the lamina propria and Peyer’s patches. This finding may open a new approach to the development of DNA vaccines for oral administration.
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