Saini Manisha, Vrati Sudhanshu
National Institute of Immunology, New Delhi 110-067, India.
J Virol. 2003 Mar;77(6):3487-94. doi: 10.1128/jvi.77.6.3487-3494.2003.
Protection against Japanese encephalitis virus (JEV) is antibody dependent, and neutralizing antibodies alone are sufficient to impart protection. Thus, we are aiming to develop a peptide-based vaccine against JEV by identifying JEV peptide sequences that could induce virus-neutralizing antibodies. Previously, we have synthesized large amounts of Johnson grass mosaic virus (JGMV) coat protein (CP) in Escherichia coli and have shown that it autoassembled to form virus-like particles (VLPs). The envelope (E) protein of JEV contains the virus-neutralization epitopes. Four peptides from different locations within JEV E protein were chosen, and these were fused to JGMV CP by recombinant DNA methods. The fusion protein autoassembled to form VLPs that could be purified by sucrose gradient centrifugation. Immunization of mice with the recombinant VLPs containing JEV peptide sequences induced anti-peptide and anti-JEV antibodies. A 27-amino-acid peptide containing amino acids 373 to 399 from JEV E protein, present on JGMV VLPs, induced virus-neutralizing antibodies. Importantly, these antibodies were obtained without the use of an adjuvant. The immunized mice showed significant protection against a lethal JEV challenge.
针对日本脑炎病毒(JEV)的保护作用依赖抗体,仅中和抗体就足以提供保护。因此,我们旨在通过鉴定可诱导病毒中和抗体的JEV肽序列来开发一种基于肽的JEV疫苗。此前,我们已在大肠杆菌中大量合成约翰逊草花叶病毒(JGMV)外壳蛋白(CP),并证明其能自动组装形成病毒样颗粒(VLP)。JEV的包膜(E)蛋白含有病毒中和表位。我们从JEV E蛋白的不同位置选择了四个肽,并通过重组DNA方法将它们与JGMV CP融合。融合蛋白自动组装形成VLP,可通过蔗糖梯度离心法纯化。用含有JEV肽序列的重组VLP免疫小鼠可诱导抗肽和抗JEV抗体。存在于JGMV VLP上的一段包含JEV E蛋白第373至399位氨基酸的27个氨基酸的肽可诱导病毒中和抗体。重要的是,这些抗体无需使用佐剂即可获得。免疫后的小鼠对致死性JEV攻击表现出显著的保护作用。