Guo Weilu, Wang Chuanhong, Song Xu, Xu Hong, Zhao Shuqing, Gu Jun, Zou Zhikun, Li Jing, Qian Jiali, Zhang Xue, Guo Rongli, Li Jizong, Li Li, Hu Zhaoyang, Ren Lili, Fan Baochao, Li Bin
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing 210014, China; School of Pharmacy, Nanjing Tech University, 5th Mofan Road, Nanjing 210009, Jiangsu, China; Taizhou Polytechnic College, Taizhou 225300, Jiangsu, China.
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Nanjing 210014, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China; School of Life Sciences, Jiangsu University, Zhenjiang 212013, China.
Vaccine. 2024 Feb 6;42(4):828-839. doi: 10.1016/j.vaccine.2024.01.020. Epub 2024 Jan 13.
Porcine epidemic diarrhea virus (PEDV) has caused serious economic losses to the pig husbandry worldwide, and the effects of existing commercialized vaccines are suboptimal. Therefore, research to develop an efficacious vaccine for prevention and control of PEDV is essential. In this study, we designed and produced trimerized proteins of full-length PEDV spike (S) protein, S1 subunit, and a tandem of multiple epitopes of S protein using an efficient mammalian expression vector system in HEK 293F cells. The immunogenicity of two commercial adjuvants, M401 and M103, was also evaluated in mice. Enzyme-linked immunosorbent assays demonstrated that all immunized mice generated highly systemic PEDV S-specific IgG and IgA antibodies. Mice in S/M103-immunized group generated the highest neutralizing antibody titer with 1:96. Compared with control group, the subunit vaccines elicited multifunctional CD3CD4 and CD3CD8 T cells, B220CD19 B cells, and CD3CD49b natural killer cells in the spleen. PEDV S/M103 vaccine, which had the best immune effect, was selected for further evaluation in piglets. Immunization with S/M103 vaccine induced high levels of S-specific IgG, IgA, and neutralizing antibodies, and increased the proliferation of peripheral blood mononuclear cells and the expression levels of interferon-γ and interleukin-4 in peripheral blood of piglets. Virus challenge test results showed significantly lower diarrheal index scores and fecal viral loads, and less pathological damage to the intestines in S/M103-immunized piglets than in controls, indicating that S/M103 provides good protection against the virulent virus challenge. Our findings suggest that trimeric PEDV S/M103 has potential as a clinical vaccine candidate.
猪流行性腹泻病毒（PEDV）已给全球养猪业造成严重经济损失，现有商业化疫苗的效果并不理想。因此，研发一种有效的PEDV预防和控制疫苗至关重要。在本研究中，我们使用高效的哺乳动物表达载体系统在HEK 293F细胞中设计并生产了全长PEDV刺突（S）蛋白、S1亚基以及S蛋白多个表位串联体的三聚体蛋白。还在小鼠中评估了两种商业佐剂M401和M103的免疫原性。酶联免疫吸附测定表明，所有免疫小鼠均产生了高度全身性的PEDV S特异性IgG和IgA抗体。S/M103免疫组小鼠产生的中和抗体效价最高，为1:96。与对照组相比，亚单位疫苗在脾脏中诱导产生了多功能CD3CD4和CD3CD8 T细胞、B220CD19 B细胞以及CD3CD49b自然杀伤细胞。选择免疫效果最佳的PEDV S/M103疫苗在仔猪中进行进一步评估。用S/M103疫苗免疫可诱导产生高水平的S特异性IgG、IgA和中和抗体，并增加仔猪外周血单个核细胞的增殖以及外周血中干扰素-γ和白细胞介素-4的表达水平。病毒攻毒试验结果显示，S/M103免疫的仔猪腹泻指数评分和粪便病毒载量显著低于对照组，肠道病理损伤也更少，表明S/M103对强毒病毒攻毒具有良好的保护作用。我们的研究结果表明，三聚体PEDV S/M103有潜力成为临床候选疫苗。
