State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Viruses. 2022 Aug 9;14(8):1746. doi: 10.3390/v14081746.
Porcine epidemic diarrhea virus (PEDV) is an enteric coronavirus that causes acute diarrhea, vomiting, dehydration, and a high mortality rate in neonatal piglets. In recent years, PEDV has been associated with co-infections with other swine enteric viruses, including porcine rotavirus (PoRV), resulting in increased mortality among newborn piglets. In this paper, we developed a bivalent vaccine against PEDV and PoRV by constructing a recombinant PEDV encoding PoRV VP7 (rPEDV-PoRV-VP7). The recombinant virus was constructed by replacing the entire open reading frame 3 (ORF3) in the genome of an attenuated PEDV strain YN150 with the PoRV VP7 gene using reverse genetic systems. Similar plaque morphology and replication kinetics were observed in Vero cells with the recombinant PEDV compared to the wild-type PEDV. It is noteworthy that the VP7 protein could be expressed stably in rPEDV-PoRV-VP7-infected cells. To evaluate the immunogenicity and safety of rPEDV-PoRV-VP7, 10-day-old piglets were vaccinated with the recombinant virus. After inoculation, no piglet displayed clinical symptoms such as vomiting, diarrhea, or anorexia. The PoRV VP7- and PEDV spike-specific IgG in serum and IgA in saliva were detected in piglets after rPEDV-PoRV-VP7 vaccination. Moreover, both PoRV and PEDV neutralizing antibodies were produced simultaneously in the inoculated piglets. Collectively, we engineered a recombinant PEDV expressing PoRV VP7 that could be used as an effective bivalent vaccine against PEDV and PoRV.
猪流行性腹泻病毒（PEDV）是一种肠道冠状病毒，可导致仔猪急性腹泻、呕吐、脱水和高死亡率。近年来，PEDV 与其他猪肠道病毒如猪轮状病毒（PoRV）的合并感染有关，导致新生仔猪死亡率增加。在本研究中，我们通过构建一种表达 PoRV VP7 的重组 PEDV（rPEDV-PoRV-VP7）来开发针对 PEDV 和 PoRV 的二价疫苗。该重组病毒是通过使用反向遗传系统，用 PoRV VP7 基因替换减毒 PEDV 株 YN150 基因组中的整个开放阅读框 3（ORF3）构建的。与野生型 PEDV 相比，重组病毒在 Vero 细胞中的蚀斑形态和复制动力学相似。值得注意的是，VP7 蛋白可以在 rPEDV-PoRV-VP7 感染的细胞中稳定表达。为了评估 rPEDV-PoRV-VP7 的免疫原性和安全性，将 10 日龄仔猪用重组病毒进行免疫接种。接种后，没有仔猪出现呕吐、腹泻或厌食等临床症状。在 rPEDV-PoRV-VP7 接种的仔猪中检测到血清中的 PoRV VP7 和 PEDV 刺突特异性 IgG 以及唾液中的 IgA。此外，接种仔猪同时产生了 PoRV 和 PEDV 中和抗体。综上所述，我们构建了一种表达 PoRV VP7 的重组 PEDV，可作为针对 PEDV 和 PoRV 的有效二价疫苗。
