核心表位区域的即插即用式纳米颗粒免疫诱导了针对PEDV的有效中和抗体和细胞免疫反应

doi:10.1016/j.jia.2024.05.002

摘要/Abstract

摘要： 目的：猪流行性腹泻病毒（PEDV）是引起猪流行性腹泻（PED）的病原体，是肠道冠状病毒的典型代表。病毒感染的有效控制取决于疫苗的不断发展。为了开发一种更新、更安全、更有效的抗PEDV亚单位疫苗，我们通过在纳米颗粒表面展示抗原来增强疫苗的免疫原性和效力。方法：利用SpyTag/SpyCatcher系统，将PEDV的核心表位结构域（COE）显示在正二十面体的mi3纳米支架上，使其自组装成含60个亚基的病毒样颗粒COE-mi3 VLPs。研究了COE-mi3 VLPs的大小、Zeta电位、微观结构和细胞毒性，并分析了它们对抗原呈递细胞摄取、抗原留存效应和树突状细胞成熟的影响。对小鼠分别进行肌肉和鼻内免疫，并分析COE-mi3 VLPs诱导的体液免疫、细胞免疫和黏膜免疫应答水平。结果：利用SpyTag/SpyCatcher的异肽键偶联效应，成功将COE展示在mi3纳米颗粒上，组装成分布均匀的COE-mi3 VLPs，并且具有良好的安全性和稳定性。COE-mi3 VLPs可以更有效地被抗原呈递细胞摄取并长久地驻留在动物体内，有助于促进树突状细胞表面标志物的表达，从而促进树突状细胞的成熟和激活。此外，COE-mi3 VLPs在小鼠模型中不仅显著改善了PEDV特异性抗体水平，还能引起更多的CD4+和CD8+ T细胞的活化和IFN-γ和IL-4细胞因子的产生。值得注意的是，COE-mi3 VLPs可以显著促进生发中心B细胞的免疫应答，对中和抗体的产生具有重要意义。此外，COE-mi3 VLPs可以通过鼻内免疫改善小鼠肠黏膜的免疫应答。结论：基于纳米颗粒多价展示的COE-mi3 VLPs可显著增强机体的体液和细胞免疫应答，通过鼻内免疫还可以提高肠道黏膜免疫水平，并且显著改善PEDV特异性中和抗体水平。综上所述，基于纳米颗粒的PEDV抗原展示具有很大的发展潜力，可以作为一种新的亚单位疫苗平台，也可能为其他肠道冠状病毒疫苗的开发提供新的思路。

Abstract: Porcine epidemic diarrhea virus (PEDV), an enteric coronavirus, is widely spread worldwide and causes huge economic losses. The effective measure to control the viral infection is to develop ideal vaccines. Here, the monomeric core epitope domain (COE) of PEDV was displayed on the surface of nanoparticles (NPs) in order to develop a newer, safer and more effective subunit vaccine against PEDV. The monomeric COE was displayed on the mi3 protein, which self-assembles into nanoparticles composed of 60 subunits, using the SpyTag/SpyCatcher system. The size, Zeta potential, microstructure of the COE-mi3 virus-like particles (VLPs)were investigated. The COE-mi3 VLPs that possessed good security, stability and better retention can be more efficiently taken up by antigen-presenting cells (APCs) and help promote dendritic cells (DCs) maturation. Moreover, COE-mi3 VLPs could prominently improve specific antibody levels including neutralizing antibodies (NAbs), and serum IgG，mucosal IgA. Moreover, COE-mi3 VLPs elicited more activation of CD4+ and CD8+ T cells and production of IFN-γ and IL-4 cytokines. In particular, COE-mi3 VLPs is an effectual antigen-delivery platform to enhance germinal center (GC) B cell responses. This structure-based self-assembly of NP gives great potential to be developed as a new subunit vaccines attractive platform, and may also provide new ideas for the development of other enteric coronavirus vaccines.

Key words: PEDV , , Nanoparticle multimerization , , Mucosal Immunization , , Germinal Center

. 核心表位区域的即插即用式纳米颗粒免疫诱导了针对PEDV的有效中和抗体和细胞免疫反应[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.05.002.

Minghui Li, Yilan Chen, Siqiao Wang, Xueke Sun, Yongkun Du, Siyuan Liu, Ruiqi Li, Zejie Chang, Peiyang Ding, Gaiping Zhang. Plug-and-display nanoparticle immunization of the core epitope domain induces potent neutralizing antibody and cellular immune responses against PEDV[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.05.002.

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