Internalin-expres sing Lactococcus lactis is able to invade small intestine of guinea pigs and deliver DNA into mammalian epithelial cells
Internalin-expres sing Lactococcus lactis is able to invade small intestine of guinea pigs and deliver DNA into mammalian epithelial cells
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DOI：
10.1016/j.micinf.2005.02.012
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发表时间：
2005-05-01
影响因子：
5.8
通讯作者：
Langella, P
中科院分区：
文献类型：
--
作者：
Guimaraes, VD;Gabriel, JE;Langella, P
使用食品级细菌乳酸乳球菌作为粘膜水平的抗原递送载体是在过去十年中深入探索的有吸引力的疫苗接种策略。在本研究中，我们开发了L。乳酸菌菌株，其可用作DNA递送载体以结合粘膜递送和DNA疫苗接种的优点。为了使乳球菌能够侵入上皮细胞，单核细胞增生李斯特菌inlA基因被克隆并在L.在天然启动子的转录控制下的乳酸。Western blot和免疫荧光分析表明，重组乳球菌有效地展示了细胞壁锚定形式的InlA。我们证明了这种表达促进了L.将Iactis inlA（+）导入人上皮细胞系Caco-2中。庆大霉素试验表明，L.乳酸杆菌在这些细胞中的表达水平比乳酸杆菌高100倍。lactis inlA（+）的活性高于野生型（wt）L.乳酸菌菌株此外，我们还证明了L.乳酸菌inlA（+）经口接种豚鼠后，能进入体内肠细胞。L. lactis inlA（+）能够将功能性真核gfp基因递送到上皮Caco-2细胞中;在1%的内化细胞中检测到GFP。洛杉矶乳酸菌inlA（+）菌株为新型口服DNA疫苗的研制提供了一种有效的载体。它也构成了一个有趣的新模型，研究在动物宿主中的细菌定位的内化的作用。(c)2005年，Elsevier SAS。All rights reserved.
The use of the food-grade bacterium Lactococcus lactis as antigen delivery vehicle at the mucosal level is an attractive vaccination strategy intensively explored during the last decade. In this study, we developed L. lactis strains which could be used as a DNA delivery vector to combine both advantages of mucosal delivery and of DNA vaccination. To render lactococci capable of invading epithelial cells, the Listeria monocytogenes inlA gene was cloned and expressed in L. lactis under transcriptional control of the native promoter. Western blot and immunofluorescence assays revealed that recombinant lactococci efficiently displayed the cell wall anchored form of InlA. We demonstrated that this expression promotes internalization of L. lactis inlA(+) into the human epithelial cell line Caco-2. Gentamicin assay showed that invasiveness of L. lactis in these cells is similar to 100-fold higher for L. lactis inlA(+) than for wild type (wt) L. lactis strains. Moreover, we showed that L. lactis inlA(+) is able to enter intestinal cells in vivo, after oral inoculation of guinea pigs. After internalization, L. lactis inlA(+) was able to deliver a functional eukaryotic gfp gene into epithelial Caco-2 cells; GFP was detected in 1% of internalized cells. The L. lactis inlA(+) strain will be a useful bacterial vector for the development of new live oral DNA vaccines. It also constitutes an interesting new model to study the role of internalin in bacterial localization in the animal host. (c) 2005 Elsevier SAS. All rights reserved.