Whittle B L, Lee E, Weir R C, Verma N K
Division of Biochemistry and Molecular Biology, Faculty of Science, Australian National University, Canberra, Australia.
J Med Microbiol. 1997 Feb;46(2):129-38. doi: 10.1099/00222615-46-2-129.
Recent developments in vaccine construction include the use of attenuated, avirulent strains of Salmonella as carriers of foreign antigens. These recombinant strains can elicit a heterologous immune response when injected into animals, demonstrating potential for their use in the construction of many vaccines. In the present study, a B-cell epitope of Murray Valley encephalitis virus (MVE) was identified and expressed in a Salmonella strain to evaluate its potential to induce a specific immune response to MVE. A synthetic oligonucleotide encoding the B-cell epitope (residues E201-224) of the envelope protein of MVE was inserted into the cloned flagellin gene of the Salmonella strain. The construct was sequenced to ensure correct orientation of the epitope. Expression of the epitope was demonstrated by Western blot analysis and immunogold electron microscopy with monoclonal antibody specific to the epitope. Electron microscopy analysis revealed multiple copies of the epitope along the flagella. The recombinant Salmonella carrying the hybrid flagellin gene elicited an immune response to the MVE epitope in a mouse model. The MVE-specific antibodies partially neutralised the virus in vitro. The significance of this system for engineering vaccines for other medically important flaviviruses is discussed.
疫苗构建的最新进展包括使用减毒、无毒的沙门氏菌菌株作为外源抗原的载体。这些重组菌株注入动物体内时可引发异源免疫反应,显示出其在多种疫苗构建中的应用潜力。在本研究中,鉴定了墨累谷脑炎病毒(MVE)的一个B细胞表位,并在一株沙门氏菌中表达,以评估其诱导针对MVE的特异性免疫反应的潜力。将编码MVE包膜蛋白B细胞表位(E201 - 224位氨基酸残基)的合成寡核苷酸插入沙门氏菌菌株的克隆鞭毛蛋白基因中。对构建体进行测序以确保表位方向正确。通过蛋白质免疫印迹分析以及使用针对该表位的单克隆抗体进行免疫金电子显微镜观察,证实了表位的表达。电子显微镜分析显示沿鞭毛有多个表位拷贝。携带杂交鞭毛蛋白基因的重组沙门氏菌在小鼠模型中引发了针对MVE表位的免疫反应。MVE特异性抗体在体外可部分中和该病毒。本文讨论了该系统对构建其他医学上重要的黄病毒疫苗的意义。