Ghiasi S M, Salmanian A H, Chinikar S, Zakeri S
Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology, Shahrak-e-Pajoohesh, 15th Km, Tehran-Karaj Highway, P.O. Box 14965-161, Tehran, Iran.
Clin Vaccine Immunol. 2011 Dec;18(12):2031-7. doi: 10.1128/CVI.05352-11. Epub 2011 Oct 19.
While Crimean-Congo hemorrhagic fever (CCHF) has a high mortality rate in humans, the associated virus (CCHFV) does not induce clinical symptoms in animals, but animals play an important role in disease transmission to humans. Our aim in this study was to examine the immunogenicity of the CCHFV glycoprotein when expressed in the root and leaf of transgenic plants via hairy roots and stable transformation of tobacco plants, respectively. After confirmatory analyses of transgenic plant lines and quantification of the expressed glycoprotein, mice were either fed with the transgenic leaves or roots, fed the transgenic plant material and injected subcutaneously with the plant-made CCHFV glycoprotein (fed/boosted), vaccinated with an attenuated CCHF vaccine (positive control), or received no treatment (negative control). All immunized groups had a consistent rise in anti-glycoprotein IgG and IgA antibodies in their serum and feces, respectively. The mice in the fed/boosted group showed a significant rise in specific IgG antibodies after a single boost. Our results imply that oral immunization of animals with edible materials from transgenic plants is feasible, and further assessments are under way. In addition, while the study of CCHF is challenging, our protocol should be further used to study CCHFV infection in the knockout mouse model and virus neutralization assays in biosafety level 4 laboratories.
虽然克里米亚-刚果出血热（CCHF）在人类中的死亡率很高，但相关病毒（CCHFV）不会在动物中引发临床症状，不过动物在疾病传播给人类的过程中起着重要作用。我们在本研究中的目的是分别通过发根和烟草植物的稳定转化来检测CCHFV糖蛋白在转基因植物的根和叶中表达时的免疫原性。在对转基因植物株系进行确证分析并对表达的糖蛋白进行定量后，给小鼠喂食转基因叶或根，喂食转基因植物材料并皮下注射植物源CCHFV糖蛋白（喂食/加强免疫），用减毒CCHF疫苗进行免疫接种（阳性对照），或不进行处理（阴性对照）。所有免疫组血清和粪便中的抗糖蛋白IgG和IgA抗体均出现持续上升。喂食/加强免疫组的小鼠在单次加强免疫后，特异性IgG抗体显著上升。我们的结果表明，用转基因植物的可食用材料对动物进行口服免疫是可行的，进一步的评估正在进行中。此外，虽然对CCHF的研究具有挑战性，但我们的方案应进一步用于在基因敲除小鼠模型中研究CCHFV感染以及在生物安全4级实验室中进行病毒中和试验。
