Rai Devendra K, Segundo Fayna Diaz-San, Schafer Elizabeth, Burrage Thomas G, Rodriguez Luis L, de Los Santos Teresa, Hoeprich Paul D, Rieder Elizabeth
Foreign Animal Disease Research Unit, United States Department of Agriculture, Agricultural Research Service, Plum Island Animal Disease Center, Greenport, NY 11944, USA; Department of Pathobiology and Veterinary Science, CANR, University of Connecticut, Storrs, CT 06269, USA.
Foreign Animal Disease Research Unit, United States Department of Agriculture, Agricultural Research Service, Plum Island Animal Disease Center, Greenport, NY 11944, USA.
Virology. 2016 Aug;495:136-47. doi: 10.1016/j.virol.2016.04.027. Epub 2016 May 20.
Here, we engineered two FMD viruses with histidine residues inserted into or fused to the FMDV capsid. Both 6xHis viruses exhibited growth kinetics, plaque morphologies and antigenic characteristics similar to wild-type virus. The 6xHis tag allowed one-step purification of the mutant virions by Co(2+) affinity columns. Electron microscopy and biochemical assays showed that the 6xHis FMDVs readily assembled into antigen: adjuvant complexes in solution, by conjugating with Ni(2+)-chelated nanolipoprotein and monophosphoryl lipid A adjuvant (MPLA:NiNLP). Animals Immunized with the inactivated 6xHis-FMDV:MPLA:NiNLP vaccine acquired enhanced protective immunity against FMDV challenge compared to virions alone. Induction of anti-6xHis and anti-FMDV neutralizing antibodies in the immunized animals could be exploited in the differentiation of vaccinated from infected animals needed for the improvement of FMD control measures. The novel marker vaccine/nanolipid technology described here has broad applications for the development of distinctive and effective immune responses to other pathogens of importance.
在此,我们构建了两种口蹄疫病毒,在口蹄疫病毒衣壳中插入或融合了组氨酸残基。两种含6xHis的病毒均表现出与野生型病毒相似的生长动力学、蚀斑形态和抗原特性。6xHis标签使得通过Co(2+)亲和柱对突变病毒粒子进行一步纯化成为可能。电子显微镜和生化分析表明,含6xHis的口蹄疫病毒通过与Ni(2+)螯合的纳米脂蛋白和单磷酰脂质A佐剂(MPLA:NiNLP)结合,能够在溶液中轻松组装成抗原:佐剂复合物。与单独的病毒粒子相比,用灭活的含6xHis的口蹄疫病毒:MPLA:NiNLP疫苗免疫的动物获得了增强的针对口蹄疫病毒攻击的保护性免疫力。在免疫动物中诱导产生的抗6xHis和抗口蹄疫病毒中和抗体可用于区分接种疫苗动物和感染动物,这对于改进口蹄疫控制措施是必要的。本文所述的新型标记疫苗/纳米脂质技术在开发针对其他重要病原体的独特且有效的免疫反应方面具有广泛应用。