Expression of Norwalk virus capsid protein in transgenic tobacco and potato and its oral immunogenicity in mice
Expression of Norwalk virus capsid protein in transgenic tobacco and potato and its oral immunogenicity in mice
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DOI：
10.1073/pnas.93.11.5335
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发表时间：
1996-05-28
影响因子：
11.1
通讯作者：
Arntzen, CJ
中科院分区：
文献类型：
--
作者：
Mason, HS;Ball, JM;Arntzen, CJ
生产重组蛋白的细胞培养系统的替代品可以以较低的成本制造非常安全的疫苗。我们已经使用基因工程植物表达候选疫苗抗原，目的是使用可食用植物器官经济地递送口服疫苗。转基因烟草和马铃薯植物被创造出来，表达诺瓦克病毒的衣壳蛋白，诺瓦克病毒是一种杯状病毒，在人类中引起流行性急性胃肠炎。从烟草叶片中提取的病毒衣壳蛋白呈38 nm的诺瓦克病毒样颗粒。重组诺瓦克病毒样颗粒（rNV）先前回收时，相同的基因在重组杆状病毒感染的昆虫细胞中表达。在烟草叶片和马铃薯块茎中表达的衣壳蛋白在蔗糖梯度中与昆虫细胞衍生的rNV共沉淀，并且在SDS/聚丙烯酰胺凝胶的免疫印迹上与昆虫细胞衍生的rNV相同。植物表达的rNV在小鼠中具有口服免疫原性。将表达rNV的烟草叶提取物通过灌胃给予CD 1小鼠，并且经处理的小鼠产生了对rNV特异的血清IgG和分泌型伊加。此外，当将表达rNV的马铃薯块茎直接喂给小鼠时，它们产生了对rNV特异的血清IgG。这些结果表明植物用于生产和递送可食用疫苗的潜在有用性。对于急需疫苗的发展中国家来说，这是一项适当的技术。
Alternatives to cell culture systems for production of recombinant proteins could make very safe vaccines at a lower cost. We have used genetically engineered plants for expression of candidate vaccine antigens with the goal of using the edible plant organs for economical delivery of oral vaccines. Transgenic tobacco and potato plants were created that express the capsid protein of Norwalk virus, a calicivirus that causes epidemic acute gastroenteritis in humans. The capsid protein could be extracted from tobacco leaves in the form of 38-nm Norwalk virus-like particles. Recombinant Norwalk virus-like particle (rNV) was previously recovered when the same gene was expressed in recombinant baculovirus-infected insect cells. The capsid protein expressed in tobacco leaves and potato tubers cosedimented in sucrose gradients with insect cell-derived rNV and appeared identical to insect cell-derived rNV on immunoblots of SDS/polyacrylamide gels. The plant-expressed rNV was orally immunogenic in mice. Extracts of tobacco leaf expressing rNV were given to CD1 mice by gavage, and the treated mice developed both serum IgG and secretory IgA specific for rNV. Furthermore, when potato tubers expressing rNV were fed directly to mice, they developed serum IgG specific for rNV. These results indicate the potential usefulness of plants for production and delivery of edible vaccines. This is an appropriate technology for developing countries where vaccines are urgently needed.