Joensuu J J, Verdonck F, Ehrström A, Peltola M, Siljander-Rasi H, Nuutila A M, Oksman-Caldentey K-M, Teeri T H, Cox E, Goddeeris B M, Niklander-Teeri V
Department of Applied Biology, P.O. Box 27, FIN-00014 University of Helsinki, Finland.
Vaccine. 2006 Mar 20;24(13):2387-94. doi: 10.1016/j.vaccine.2005.11.056. Epub 2005 Dec 9.
Transgenic plants are attractive bioreactors to large-scale production of recombinant proteins because of their relatively low cost. This study reports for the first time the use of transgenic plants to reduce enterotoxigenic Escherichia coli (ETEC) excretion in its natural host species. The DNA sequence encoding the major subunit and adhesin FaeG of F4+ ETEC was transformed into edible alfalfa plants. Targeting of FaeG production to chloroplasts led to FaeG levels of up to 1% of the total soluble protein fraction of the transgenic alfalfa. Recombinant plant-produced FaeG (pFaeG) remained stable for 2 years when the plant material was dried and stored at room temperature. Intragastric immunization of piglets with pFaeG induced a weak F4-specific humoral response. Co-administration of pFaeG and the mucosal adjuvant cholera toxin (CT) enhanced the immune response against FaeG, reflected a better induction of an F4-specific immune response. In addition, the intragastric co-administration of CT with pFaeG significantly reduced F4+ E. coli excretion following F4+ ETEC challenge as compared with pigs that had received nontransgenic plant material. In conclusion, transgenic plants producing the FaeG subunit protein could be used for production and delivery of oral vaccines against F4+ ETEC infections.
由于成本相对较低,转基因植物是大规模生产重组蛋白的理想生物反应器。本研究首次报道了利用转基因植物减少天然宿主物种中产肠毒素大肠杆菌(ETEC)的排泄。将编码F4+ ETEC主要亚基和黏附素FaeG的DNA序列转化到可食用的苜蓿植物中。将FaeG的生产靶向叶绿体,使得转基因苜蓿中FaeG的水平高达总可溶性蛋白组分的1%。当植物材料干燥并在室温下储存时,重组植物产生的FaeG(pFaeG)可保持稳定2年。用pFaeG对仔猪进行胃内免疫诱导了较弱的F4特异性体液反应。pFaeG与黏膜佐剂霍乱毒素(CT)共同给药增强了针对FaeG的免疫反应,反映出对F4特异性免疫反应的更好诱导。此外,与接受非转基因植物材料的猪相比,CT与pFaeG胃内共同给药显著减少了F4+ ETEC攻击后F4+大肠杆菌的排泄。总之,产生FaeG亚基蛋白的转基因植物可用于生产和递送针对F4+ ETEC感染的口服疫苗。