Blokhina Elena A, Mardanova Eugenia S, Zykova Anna A, Stepanova Liudmila A, Shuklina Marina A, Tsybalova Liudmila M, Ravin Nikolai V
Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia.
Smorodintsev Research Institute of Influenza, Russian Ministry of Health, 197376 St. Petersburg, Russia.
Plants (Basel). 2023 Jun 5;12(11):2228. doi: 10.3390/plants12112228.
Despite advances in vaccine development, influenza remains a persistent global health threat and the search for a broad-spectrum recombinant vaccine against influenza continues. The extracellular domain of the transmembrane protein M2 (M2e) of the influenza A virus is highly conserved and can be used to develop a universal vaccine. M2e is a poor immunogen by itself, but it becomes highly immunogenic when linked to an appropriate carrier. Here, we report the transient expression of a recombinant protein comprising four tandem copies of M2e fused to an artificial self-assembling peptide (SAP) in plants. The hybrid protein was efficiently expressed in using the self-replicating potato virus X-based vector pEff. The protein was purified using metal affinity chromatography under denaturing conditions. The hybrid protein was capable of self-assembly in vitro into spherical particles 15-30 nm in size. The subcutaneous immunization of mice with M2e-carrying nanoparticles induced high levels of M2e-specific IgG antibodies in serum and mucosal secretions. Immunization provided mice with protection against a lethal influenza A virus challenge. SAP-based nanoparticles displaying M2e peptides can be further used to develop a recombinant "universal" vaccine against influenza A produced in plants.
尽管疫苗研发取得了进展，但流感仍然是一个持续存在的全球健康威胁，针对流感的广谱重组疫苗的研发仍在继续。甲型流感病毒跨膜蛋白M2（M2e）的胞外结构域高度保守，可用于开发通用疫苗。M2e本身是一种弱免疫原，但与合适的载体连接后会变得具有高度免疫原性。在此，我们报道了一种重组蛋白在植物中的瞬时表达，该重组蛋白由四个串联的M2e拷贝与一种人工自组装肽（SAP）融合而成。使用基于马铃薯X病毒的自我复制载体pEff在植物中高效表达了该杂合蛋白。在变性条件下通过金属亲和层析纯化该蛋白。该杂合蛋白能够在体外自组装成大小为15 - 30纳米的球形颗粒。用携带M2e的纳米颗粒对小鼠进行皮下免疫可诱导血清和粘膜分泌物中产生高水平的M2e特异性IgG抗体。免疫为小鼠提供了针对致死性甲型流感病毒攻击的保护。展示M2e肽的基于SAP的纳米颗粒可进一步用于开发在植物中生产的针对甲型流感的重组“通用”疫苗。
