PDF(Pubmed)
Abstract：
As a highly pathogenic avian virus, H5 influenza poses a serious threat to livestock, the poultry industry, and public health security. Hemagglutinin (HA) is both the dominant epitope and the main target of influenza-neutralizing antibodies. Here, we designed a nanoparticle hemagglutinin influenza vaccine to improve the immunogenicity of the influenza vaccine. In this study, HA5 subtype influenza virus was used as the candidate antigen and was combined with the artificially designed double-branch scaffold protein I53_dn5 A and B. A structurally correct and bioactive trimer HA5-I53_dn5B/Y98F was obtained through secretion and purification using an insect baculovirus expression system; I53_dn5A was obtained by purification using a prokaryotic expression system. HA5-I53_dn5B/Y98F and I53_dn5A self-assembled into spherical nanoparticles (HA5-I53_dn5) in vitro with a diameter of about 45 nm. Immunization and serum test results showed that both HA5-I53_dn5B/Y98F and HA5-I53_dn5 could induce HA5-specific antibodies; however, the immunogenicity of HA5-I53_dn5 was better than that of HA5-I53_dn5B/Y98F. Groups treated with HA5-I53_dn5B and HA5-I53_dn5 nanoparticles produced IgG antibody titers that were not statistically different from those of the nanoparticle-containing adjuvant group. This production of trimerized HA5-I53_dn5B and HA5-I53_dn5 nanoparticles using baculovirus expression provides a reference for the development of novel, safe, and efficient influenza vaccines.
摘要：
作为一种高致病性禽流感病毒，H5流感对牲畜构成严重威胁,家禽业,和公共卫生安全。血凝素(HA)是流感中和抗体的优势表位和主要靶标。这里,我们设计了一种纳米血凝素流感疫苗来提高流感疫苗的免疫原性.在这项研究中,以HA5亚型流感病毒为候选抗原，与人工设计的双分支支架蛋白I53_dn5A和B结合，利用昆虫杆状病毒表达系统通过分泌纯化获得结构正确、具有生物活性的三聚体HA5-I53_dn5B/Y98F；利用原核表达系统纯化获得I53_dn5A。HA5-I53_dn5B/Y98F和I53_dn5A在体外自组装成球形纳米颗粒(HA5-I53_dn5)，直径约为45nm。免疫和血清检测结果表明,HA5-I53_dn5B/Y98F和HA5-I53_dn5均能诱导HA5特异性抗体;HA5-I53_dn5的免疫原性优于HA5-I53_dn5B/Y98F。用HA5-I53_dn5B和HA5-I53_dn5纳米颗粒处理的组产生的IgG抗体滴度与含有纳米颗粒的佐剂组的IgG抗体滴度没有统计学差异。这种利用杆状病毒表达的三聚HA5-I53_dn5B和HA5-I53_dn5纳米粒的生产为开发新型,安全,和有效的流感疫苗。
