Tsybalova Liudmila M, Stepanova Liudmila A, Kuprianov Victor V, Blokhina Elena A, Potapchuk Marina V, Korotkov Alexander V, Gorshkov Andrey N, Kasyanenko Marina A, Ravin Nikolai V, Kiselev Oleg I
Research Institute of Influenza, Russian Federation Ministry of Health, St. Petersburg, Russia.
Research Institute of Influenza, Russian Federation Ministry of Health, St. Petersburg, Russia.
Vaccine. 2015 Jun 26;33(29):3398-406. doi: 10.1016/j.vaccine.2015.04.073. Epub 2015 May 11.
A long-term objective when designing influenza vaccines is to create one with broad cross-reactivity that will provide effective control over influenza, no matter which strain has caused the disease. Here we summarize the results from an investigation into the immunogenic and protective capacities inherent in variations of a recombinant protein, HBc/4M2e. This protein contains four copies of the ectodomain from the influenza virus protein M2 (M2e) fused within the immunodominant loop of the hepatitis B virus core antigen (HBc). Variations of this basic design include preparations containing M2e from the consensus human influenza virus; the M2e from the highly pathogenic avian A/H5N1 virus and a combination of two copies from human and two copies from avian influenza viruses. Intramuscular delivery in mice with preparations containing four identical copies of M2e induced high IgG titers in blood sera and bronchoalveolar lavages. It also provoked the formation of memory T-cells and antibodies were retained in the blood sera for a significant period of time post immunization. Furthermore, these preparations prevented the death of 75-100% of animals, which were challenged with lethal doses of virus. This resulted in a 1.2-3.5 log10 decrease in viral replication within the lungs. Moreover, HBc particles carrying only "human" or "avian" M2e displayed cross-reactivity in relation to human (A/H1N1, A/H2N2 and A/H3N2) or A/H5N1 and A(H1N1)pdm09 viruses, respectively; however, with the particles carrying both "human" and "avian" M2e this effect was much weaker, especially in relation to influenza virus A/H5N1. It is apparent from this work that to quickly produce vaccine for a pandemic it would be necessary to have several variations of a recombinant protein, containing four copies of M2e (each one against a group of likely influenza virus strains) with these relevant constructs housed within a comprehensive collection Escherichia coli-producers and maintained ready for use.
设计流感疫苗的一个长期目标是研发出一种具有广泛交叉反应性的疫苗，无论引发疾病的是哪种毒株，它都能有效控制流感。在此，我们总结了对重组蛋白HBc/4M2e变体的免疫原性和保护能力进行调查的结果。该蛋白包含来自流感病毒蛋白M2（M2e）胞外域的四个拷贝，融合于乙肝病毒核心抗原（HBc）的免疫显性环内。这种基本设计的变体包括含有人流感病毒共有序列的M2e制剂；高致病性禽流感A/H5N1病毒的M2e，以及来自人流感病毒的两个拷贝和禽流感病毒的两个拷贝的组合。用含有四个相同拷贝M2e的制剂对小鼠进行肌肉注射，可在血清和支气管肺泡灌洗液中诱导产生高IgG滴度。它还促使记忆T细胞形成，并且在免疫后相当长一段时间内抗体仍保留在血清中。此外，这些制剂可防止75%至100%接受致死剂量病毒攻击的动物死亡。这导致肺部病毒复制减少1.2至3.5个对数10。此外，仅携带“人源”或“禽源”M2e的HBc颗粒分别对人（A/H1N1、A/H2N2和A/H3N2）或A/H5N1及A(H1N1)pdm09病毒表现出交叉反应性；然而，携带“人源”和“禽源”M2e的颗粒这种效应要弱得多，尤其是相对于A/H5N1流感病毒。从这项工作可以明显看出，为了快速生产大流行疫苗，有必要拥有几种重组蛋白变体，其包含四个拷贝的M2e（每个拷贝针对一组可能的流感病毒毒株），这些相关构建体保存在一组全面的大肠杆菌生产菌株中并随时可供使用。
