Mason H S, Haq T A, Clements J D, Arntzen C J
Boyce Thompson Institute for Plant Research, Ithaca, NY 14853-1801, USA.
Vaccine. 1998 Aug;16(13):1336-43. doi: 10.1016/s0264-410x(98)80020-0.
The authors have designed and constructed a plant-optimize synthetic gene encoding the Escherichia coli heat-labile enterotoxin B subunit (LT-B), for use in transgenic plants as an edible vaccine against enterotoxigenic E. coli. Expression of the synthetic LT-B gene in potato plants under the control of a constitutive promoter yielded increased accumulation of LT-B in leaves and tubers, as compared to the bacterial LT-B gene. The plant-derived LT-B assembled into native pentameric structures as evidenced by its ability to bind ganglioside. The authors demonstrated immunogenicity by feeding mice the raw tubers and comparing the anti-LT-B serum IgG and faecal IgA to that produced in mice gavaged with bacterial LT-B. Mice were fed three weekly doses of 5 g tuber tissue containing either 20 or 50 micrograms LT-B, or gavaged weekly with 5 micrograms of LT-B from recombinant E. coli. One week after the third dose, mice immunized with potato LT-B had higher levels of serum and mucosal anti-LT-B than those gavaged with bacterial LT-B. Mice were challenged by oral administration of 25 micrograms LT, and protection assessed by comparing the gut/carcass mass ratios. Although none of the mice were completely protected, the higher dose potato vaccine compared favourably with the bacterial vaccine. These findings show that an edible vaccine against E. coli LT-B is feasible.
作者设计并构建了一种经过植物优化的合成基因,该基因编码大肠杆菌不耐热肠毒素B亚基(LT-B),用于转基因植物,作为针对产肠毒素大肠杆菌的可食用疫苗。与细菌LT-B基因相比,在组成型启动子控制下,合成LT-B基因在马铃薯植株中的表达使叶片和块茎中LT-B的积累量增加。植物来源的LT-B组装成天然五聚体结构,这可通过其结合神经节苷脂的能力得到证明。作者通过给小鼠喂食生块茎,并将抗LT-B血清IgG和粪便IgA与用细菌LT-B灌胃的小鼠所产生的进行比较,证明了其免疫原性。给小鼠每周喂食三次剂量为5克的块茎组织,其中含有20或50微克LT-B,或每周用5微克重组大肠杆菌来源的LT-B进行灌胃。在第三次给药后一周,用马铃薯LT-B免疫的小鼠血清和黏膜抗LT-B水平高于用细菌LT-B灌胃的小鼠。通过口服给予25微克LT对小鼠进行攻毒,并通过比较肠道/胴体质量比来评估保护效果。尽管没有小鼠得到完全保护,但高剂量的马铃薯疫苗与细菌疫苗相比具有优势。这些发现表明,针对大肠杆菌LT-B的可食用疫苗是可行的。