Immunogenicity of a plant-derived edible chimeric EspA, Intimin and Tir of Escherichia coli O157:H7 in mice
Immunogenicity of a plant-derived edible chimeric EspA, Intimin and Tir of Escherichia coli O157:H7 in mice
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DOI：
10.1016/j.plantsci.2011.01.004
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发表时间：
2011-04-01
期刊：
影响因子：
5.2
通讯作者：
Salmanian, Ali Hatef
中科院分区：
文献类型：
--
作者：
Amani, Jafar;Mousavi, Seyed Latif;Salmanian, Ali Hatef
转基因植物提供了以低生产成本和最小纯化或富集大规模生产和递送口服免疫原的可能性。牛是大肠杆菌O157：H7的重要宿主，在动物中开发特异性免疫力将是非常宝贵的。肠出血性大肠杆菌LEE基因座表达的毒力因子有Intimin、Tir和EspA蛋白。杆菌我们假设这些效应物的嵌合重组形式作为可食用疫苗递送将减少小鼠中细菌的定殖。构建了由espA（e）、eae（i）和tir（t）通过连接子连接而成的合成基因（eit）。将该基因进行密码子优化，并克隆到与CaMV 35S和FAE启动子相邻的植物表达载体中，用于在烟草和卡诺拉植物中表达。转基因烟草叶片和油菜籽中可溶性蛋白含量分别为0.2%和0.3%。用重组EIT皮下或口服免疫小鼠，并用E. coli 0157：H7的细菌脱落显著减少。利用转基因植物作为免疫原是防治大肠杆菌病的有效手段。coli O157：H7。(C)2011爱思唯尔爱尔兰有限公司保留所有权利。
Transgenic plants offer the possibility to produce and deliver an oral immunogen on a large-scale with low production costs and minimal purification or enrichment. Cattles are important reservoirs of Escherichia coli O157:H7 and developing a specific immunity in animals would be invaluable. Intimin, Tir, and EspA proteins are the virulence factors expressed by LEE locus of enterohemorrhagic E. coli. We hypothesized that the chimeric recombinant forms of these effectors delivered as an edible-base vaccine would reduce colonization of bacteria in mice. A synthetic gene (eit) composed of espA (e), eae (i) and tir (t) attached by linkers was constructed. The gene was codon optimized and cloned into plant expression vectors adjacent to CaMV35S and FAE promoters for expression in tobacco and canola plants. Of total soluble protein 0.2% and 0.3% (in average) were detected in transgenic tobacco leaves and canola seeds respectively. Mice immunized either subcutaneously or orally with recombinant EIT and challenged with E. coli 0157:H7 significantly exhibited reduced bacterial shedding. Application of transgenic plants containing trivalent immunogen is an effective tool for protection against E. coli O157:H7. (C) 2011 Elsevier Ireland Ltd. All rights reserved.