Dual-linker gold nanoparticles as adjuvanting carriers for multivalent display of recombinant influenza hemagglutinin trimers and flagellin improve the immunological responses in vivo and in vitro.
Dual-linker gold nanoparticles as adjuvanting carriers for multivalent display of recombinant influenza hemagglutinin trimers and flagellin improve the immunological responses in vivo and in vitro.
复制标题
DOI：
10.2147/ijn.s137222
复制
发表时间：
2017
影响因子：
8
通讯作者：
Wang BZ
中科院分区：
文献类型：
--
作者：
Wang C;Zhu W;Wang BZ
接种疫苗是控制传染病的最具成本效益的手段。尽管目前的流感疫苗在对抗紧密匹配的毒株方面是有效的，但是这样的疫苗具有主要的局限性，例如需要每个季节生产新的疫苗、依赖于鸡蛋的生产系统、长的生产周期、将疫苗与流行毒株匹配的不确定性、以及不能对由基因漂移或转变引起的新的流感大流行作出反应。为了克服传统流感疫苗的内在局限性，我们设计了与重组三重A/Aichi/2/68（H3 N2）、血凝素（HA）和TLR 5激动剂鞭毛蛋白（FliC）缀合的双接头金纳米颗粒（AuNPs）作为新的疫苗方法。使用点击化学和金属螯合反应来偶联这两种蛋白质。发现缀合的蛋白质具有高偶联特异性、在恶劣环境中的高稳定性、高缀合效率以及保持用于免疫原性和免疫刺激的适当蛋白质构象的能力。当经由小鼠中的单次鼻内免疫施用时，AuNPs-HA/FliC和AuNPs-HA制剂均诱导比可溶性HA和FliC蛋白的混合物更高水平的抗体应答。为了进一步研究这些纳米颗粒的亲和力，在JAWS II树突状细胞（DC）系和骨髓来源的DC（BMDC）模型中进行体外实验。结果表明，双缀合的AuNPs被DC快速靶向并摄取。因此，DC被诱导成熟，如高水平的细胞因子分泌和膜共刺激分子表达所证明的。当脾T细胞与AuNPs-HA/FliC致敏的BMDC共培养时，观察到T细胞增殖。这些结果表明，双缀合的AuNP在以定向的多价形式同时展示抗原和佐剂方面是有效的，并且可以通过激活DC和T细胞来促进强烈的免疫应答。
Vaccination is the most cost-effective means of infectious disease control. Although current influenza vaccines are effective in battling closely matched strains, such vaccines have major limitations such as the requirement to produce new vaccines every season, an egg-dependent production system, long production periods, uncertainty in matching the vaccine to circulating strains, and the inability to react to new influenza pandemics resulting from genetic drift or shift. To overcome the intrinsic limitations of the conventional influenza vaccine, we have designed dual-linker gold nanoparticles (AuNPs) conjugated with both recombinant trimetric A/Aichi/2/68 (H3N2), hemagglutinin (HA) and TLR5 agonist flagellin (FliC) as a novel vaccine approach. Click chemistry and metal-chelating reactions were used to couple the two proteins. The conjugated proteins were found to possess high coupling specificity, high stability in harsh environments, high conjugation efficiency, and the ability to keep the appropriate protein conformations for immunogenicity and immunostimulation. Both AuNPs-HA/FliC and AuNPs-HA formulations induced higher levels of antibody responses than a mixture of soluble HA and FliC proteins when administered via a single intranasal immunization in mice. To further investigate the adjuvancy of these nanoparticles, in vitro experiments were conducted in both the JAWS II dendritic cell (DC) line and bone marrow-derived DC (BMDC) models. The results showed that dual-conjugated AuNPs were rapidly targeted and taken up by DCs. Consequently, DCs were induced toward maturation, as demonstrated by high levels of cytokine secretions and membrane costimulatory molecule expression. T cell proliferation was observed when splenic T cells were cocultured with AuNPs-HA/FliC-primed BMDCs. These results suggest that dual-conjugated AuNPs are effective at simultaneously displaying antigens and adjuvants in an oriented, multivalent format and can promote a strong immune response by activating DCs and T cells.