Expression and Characterization of Recombinant, Tetrameric and Enzymatically Active Influenza Neuraminidase for the Setup of an Enzyme-Linked Lectin-Based Assay
Expression and Characterization of Recombinant, Tetrameric and Enzymatically Active Influenza Neuraminidase for the Setup of an Enzyme-Linked Lectin-Based Assay
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DOI：
10.1371/journal.pone.0135474
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发表时间：
2015-08-17
期刊：
影响因子：
3.7
通讯作者：
Cozzi, Roberta
中科院分区：
文献类型：
--
作者：
Prevato, Marua;Ferlenghi, Ilaria;Cozzi, Roberta
开发诱导广谱、长期免疫的通用流感疫苗已成为流感疫苗研发的一个重要的潜在可实现目标。血凝素（HA）和神经氨酸酶（NA）是两种主要的流感病毒抗原。尽管针对流感病毒的抗体反应主要针对 HA，但据报道 NA 的遗传稳定性更高；因此，基于 NA 的疫苗有可能在较长时间内有效。 NA特异性免疫已被证明可以限制流感病毒的传播，从而减轻疾病症状，并在小鼠攻击实验中提供针对异亚型病毒的交叉保护。大量高纯度和稳定的NA的生产可能有利于新型抗病毒药物、基于亚单位的疫苗和新型诊断工具的开发。在这项研究中，在哺乳动物细胞中从猪 A/California/07/2009 (H1N1) 和禽 A/turkey/Turkey/01/2005 (H5N1) 流感病毒中产生高水平的重组 NA (rNA)。生化、结构和免疫学特征表明，产生的可溶性 rNA 是四聚体，具有酶活性和免疫原性，最终它们代表了酶联凝集素测定 (ELLA) 中常规使用的 NA 来源的良好替代品。
Developing a universal influenza vaccine that induces broad spectrum and longer-term immunity has become an important potentially achievable target in influenza vaccine research and development. Hemagglutinin (HA) and neuraminidase (NA) are the two major influenza virus antigens. Although antibody responses against influenza virus are mainly directed toward HA, NA is reported to be more genetically stable; hence NA-based vaccines have the potential to be effective for longer time periods. NA-specific immunity has been shown to limit the spread of influenza virus, thus reducing disease symptoms and providing cross-protection against heterosubtypic viruses in mouse challenge experiments.The production of large quantities of highly pure and stable NA could be beneficial for the development of new antivirals, subunit-based vaccines, and novel diagnostic tools. In this study, recombinant NA (rNA) was produced in mammalian cells at high levels from both swine A/California/07/2009 (H1N1) and avian A/turkey/Turkey/01/2005 (H5N1) influenza viruses. Biochemical, structural, and immunological characterizations revealed that the soluble rNAs produced are tetrameric, enzymatically active and immunogenic, and finally they represent good alternatives to conventionally used sources of NA in the Enzyme-Linked Lectin Assay (ELLA).