Virus-like particle vaccines with epitopes from porcine epidemic virus and transmissible gastroenteritis virus incorporated into self-assembling ADDomer platform provide clinical immune responses in piglets.

Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) are major intestinal coronaviruses that cause vomiting, diarrhea, dehydration, and mortality in piglets. These viruses coexist and lead to significant economic losses in the swine industry. Virus-like particles (VLPs) have emerged as promising alternatives to conventional inactivated vaccines due to their exceptional safety, efficacy, and ability to provide multi-disease protection with a single dose. Our study focused on specific antigenic epitopes from the PEDV S protein (SS2 and 2C10 regions) and the TGEV S protein (A and D sites) as target candidates. These epitopes were integrated into the ADDomer framework, and we successfully generated recombinant proteins AD, AD-P, AD-T, and AD-PT using the baculovirus expression vector system (BEVS). By meticulously optimizing conditions in High Five cells, we successfully expressed and purified the recombinant proteins. Subsequently, we developed the recombinant ADDomer-VLP vaccine and conducted a comprehensive evaluation of its efficacy in piglets. Following ultrafiltration concentration and sucrose gradient centrifugation purification, the recombinant proteins self-assembled into VLPs as observed by transmission electron microscopy (TEM). Administration of the vaccine did not result in any adverse reactions in the immunized piglets. Additionally, no significant instances of fever were detected in any of the experimental groups, and there were no notable changes in average daily weight gain compared to the control group that received PBS. The recombinant ADDomer-VLP vaccines demonstrated strong immunogenicity, effectively stimulating the production of neutralizing antibodies against both PEDV and TGEV. Moreover, the recombinant ADDomer-VLP vaccine induced elevated levels of IFN-γ, IL-2, and IL-4, and enhanced cytotoxic T lymphocyte (CTL) activity in the peripheral blood of piglets. These recombinant VLPs have demonstrated the ability to induce strong cellular and humoral immune responses in piglets, making them an incredibly promising platform for the rapid and simplified development of epitope vaccines.

猪的流行性腹泻病毒（PEDV）和可传播的胃炎病毒（TGEV）是引起呕吐，腹泻，脱水和死亡率的主要肠道冠状病毒。 VLP）由于其出色的安全性，效率和提供单剂量提供多疾病保护的能力，因此已成为常规灭活疫苗的承诺替代品。 我们的研究重点是来自PEDV S蛋白（SS2和2C10区）的特定抗原表位和TGEV的蛋白（A和D位点）作为靶标的候选物。 AD-P，AD-T和AD-PT使用杆状病毒表达载体系统（BEV），通过精心优化高五个细胞的条件，我们成功表达并纯化了重组蛋白。并对其在小猪中的有效性进行了全面的评估。 在超滤浓度和蔗糖梯度离心纯化之后，通过透射电子显微镜（TEM）观察到的重组蛋白自组装成VLP。在任何实验组中都检测到它们，与接受PBS的对照组相比，每日平均体重增加没有明显变化。此外，TGEV。重组addomer-VLP疫苗诱导了IFN-γ，IL-2和IL-4的水平，并增强了小猪外围血液中的细胞毒性T淋巴细胞（CTL）活性。 这些重组VLP已证明能够在小猪中诱导强烈的细胞和体液免疫调查，这使它们成为了表位疫苗快速而简化开发的令人难以置信的有望平台。