Engineering and evaluation of amyloid assemblies as a nanovaccine against the Chikungunya virus
Engineering and evaluation of amyloid assemblies as a nanovaccine against the Chikungunya virus
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DOI：
10.1039/c8nr05948a
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发表时间：
2018-11-07
期刊：
影响因子：
6.7
通讯作者：
Bourgault, Steve
中科院分区：
文献类型：
--
作者：
Babych, Margaryta;Bertheau-Mailhot, Genevieve;Bourgault, Steve
暴露高密度抗原的纳米颗粒的设计构成了解决常规减毒活疫苗的安全性问题以及增加亚单位疫苗的免疫原性的有希望的策略。在这项研究中，我们开发了一种完全合成的纳米疫苗的基础上，淀粉样蛋白肽序列具有高自组装性能。来自基孔肯雅病毒的E2糖蛋白的免疫原性表位E2 EP 3被用于评估源自内源性淀粉样蛋白生成多肽的10-mer肽作为新型疫苗平台的潜力。通过固相合成制备嵌合肽，其包含通过短的柔性接头连接至淀粉样蛋白核心的肽抗原。如使用原子力显微镜观察到的，这些多肽自组装成直径范围为6至8 nm的线性和无分支的原纤维。一个四元构象丰富的交叉片，其特征在于这些组件，证明了圆二色光谱和硫磺素T荧光。免疫金标记的纤维的ELISA测定和透射电子显微镜显示高密度的基孔肯雅病毒E2糖蛋白衍生的表位暴露在纤维表面上。这些淀粉样纤维是细胞相容的，并被巨噬细胞有效地摄取。小鼠免疫显示针对E2 EP 3表位的强IgG应答，其依赖于自组装并且不需要共注射Alhydrogel佐剂。这些结果表明，交叉折叠淀粉样蛋白组装体构成合适的合成自佐剂组装体以锚抗原决定簇并增加肽表位的免疫原性。
The design of nanoparticles exposing a high density of antigens constitutes a promising strategy to address safety concerns of conventional life-attenuated vaccines as well as to increase the immunogenicity of subunit vaccines. In this study, we developed a fully synthetic nanovaccine based on an amyloid peptide sequence with high self-assembling properties. The immunogenic epitope E2EP3 from the E2 glycoprotein of the Chikungunya virus was used to evaluate the potential of a 10-mer peptide derived from an endogenous amyloidogenic polypeptide as a novel vaccine platform. Chimeric peptides, comprising the peptide antigen attached to the amyloid core by a short flexible linker, were prepared by solid phase synthesis. As observed using atomic force microscopy, these polypeptides self-assembled into linear and unbranched fibrils with a diameter ranging from 6 to 8 nm. A quaternary conformation rich in cross--sheets characterized these assemblies, as demonstrated by circular dichroism spectroscopy and thioflavin T fluorescence. ELISA assays and transmission electronic microscopy of immunogold labeled-fibrils revealed a high density of the Chikungunya virus E2 glycoprotein derived epitope exposed on the fibril surface. These amyloid fibrils were cytocompatible and were efficiently uptaken by macrophages. Mice immunization revealed a robust IgG response against the E2EP3 epitope, which was dependent on self-assembly and did not require co-injection of the Alhydrogel adjuvant. These results indicate that cross--sheet amyloid assemblies constitute suitable synthetic self-adjuvanted assemblies to anchor antigenic determinants and to increase the immunogenicity of peptide epitopes.