CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
Protein Cell. 2011 Dec;2(12):997-1005. doi: 10.1007/s13238-011-1134-y. Epub 2012 Jan 10.
There is a great need for new vaccine development against influenza A viruses due to the drawbacks of traditional vaccines that are mainly prepared using embryonated eggs. The main component of the current split influenza A virus vaccine is viral hemagglutinin (HA) which induces a strong antibody-mediated immune response. To develop a modern vaccine against influenza A viruses, the current research has been focused on the universal vaccines targeting viral M2, NP and HA proteins. Crystallographic studies have shown that HA forms a trimer embedded on the viral envelope surface, and each monomer consists of a globular head (HA1) and a "rod-like" stalk region (HA2), the latter being more conserved among different HA subtypes and being the primary target for universal vaccines. In this study, we rationally designed the HA head based on the crystal structure of the 2009-pandemic influenza A (H1N1) virus HA as a model, tested its immunogenicity in mice, solved its crystal structure and further examined its immunological characteristics. The results show that the HA globular head can be easily prepared by in vitro refolding in an E. coli expression system, which maintains its intact structure and allows for the stimulation of a strong immune response. Together with recent reports on some similar HA globular head preparations we conclude that structure-based rational design of the HA globular head can be used for subtype-specific vaccines against influenza viruses.
由于传统疫苗主要使用鸡胚制备,存在诸多缺点,因此非常需要开发针对甲型流感病毒的新型疫苗。目前,分裂型甲型流感病毒疫苗的主要成分是病毒血凝素 (HA),它能诱导强烈的抗体介导的免疫反应。为了开发针对甲型流感病毒的现代疫苗,目前的研究集中在针对病毒 M2、NP 和 HA 蛋白的通用疫苗上。晶体学研究表明,HA 形成三聚体嵌入在病毒包膜表面,每个单体由球形头部(HA1)和“棒状”茎部区域(HA2)组成,后者在不同的 HA 亚型之间更保守,是通用疫苗的主要靶点。在这项研究中,我们基于 2009 年大流行的甲型流感(H1N1)病毒 HA 的晶体结构,对 HA 头部进行了合理设计,作为模型,在小鼠中测试了其免疫原性,解析了其晶体结构,并进一步研究了其免疫学特性。结果表明,HA 球形头部可以通过在大肠杆菌表达系统中进行体外重折叠轻松制备,保持其完整的结构,并能刺激强烈的免疫反应。结合最近关于一些类似的 HA 球形头部制剂的报道,我们得出结论,基于结构的 HA 球形头部的合理设计可用于针对流感病毒的亚型特异性疫苗。