College of Animal Science and Technology, Hainan Key Lab of Tropical Animal Reproduction and Breeding and Epidemic Disease Research, Hainan University, Haidian Island, Haikou, 570228, China.
College of Animal Science & Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, 163319, Heilongjiang, China.
Appl Microbiol Biotechnol. 2020 Apr;104(7):3011-3023. doi: 10.1007/s00253-020-10420-6. Epub 2020 Jan 31.
Bovine herpesvirus type 1 (BoHV-1) causes considerable economic losses to the cow industry. Vaccination remains an effective strategy to control the diseases associated with BoHV-1. However, live vaccines present safety concerns, especially in pregnant cows; thus, nonreplicating vaccines have been developed to control the disease. The envelope glycoproteins of BoHV-1 induce a protective immune response. In this work, selected epitopes on glycoproteins gD, gC, and gB were constructed in triplicate with linker peptides. Vaccination of rabbits demonstrated that P2-gD/gC/gB with AAYAAY induced higher specific antibodies than that with GGGGS linker. P2-gD/gC/gB with AAYAAY linker was fused with bovine interleukin-6 (BoIL-6) or rabbit IL-6 (RaIL-6) and bacterially expressed. Rabbits were intramuscularly immunized with 100 μg of P2-gD/gC/gB-BoIL-6, P2-gD/gC/gB-RaIL-6, P2-gD/gC/gB, P2-gD/gC/gB plus BoIL-6, P2-(gD-a)3-BoIL-6, or P2-(gD-a)3 emulsified with ISA 206 adjuvant thrice at 3-week intervals. P2-gD/gC/gB-BoIL-6 generated a higher titer of BoHV-1-specific antibodies, neutralizing antibodies, interferon (IFN)-γ, and IL-4 compared with P2-gD/gC/gB plus BoIL-6, P2-gD/gC/gB-RaIL-6, or other formulation. P2-gD/gC/gB-BoIL-6 triggered similar levels of antibodies and significantly higher titer of IFN-γ and IL-4 compared with inactivated bovine viral diarrhea (BVD)-infectious bovine rhinotracheitis (IBR) vaccine. Rabbits vaccinated with P2-gD/gC/gB-BoIL-6 dramatically reduced viral shedding and tissue lesions in lungs and trachea after viral challenge and reactivation compared with those with P2-gD/gC/gB plus BoIL-6 or P2-gD/gC/gB-RaIL-6. P2-gD/gC/gB-BoIL-6 provided protective effects against viral shedding and tissue pathogenesis similar to those of the inactivated vaccine. The data confirmed the safety and immunogenicity of multiple-epitope recombinant protein and a potential vaccine candidate to control the disease, especially for pregnant cattle.
牛疱疹病毒 1 型（BoHV-1）给奶牛业造成了巨大的经济损失。疫苗接种仍然是控制与 BoHV-1 相关疾病的有效策略。然而，活疫苗存在安全隐患，特别是在怀孕的奶牛中；因此，已经开发出非复制疫苗来控制这种疾病。BoHV-1 的包膜糖蛋白诱导保护性免疫应答。在这项工作中，用连接肽重复构建了糖蛋白 gD、gC 和 gB 上的选定表位。对兔子进行的疫苗接种表明，与 GGGGS 接头相比，具有 AAYAAY 的 P2-gD/gC/gB 诱导了更高的特异性抗体。具有 AAYAAY 接头的 P2-gD/gC/gB 与牛白细胞介素-6（BoIL-6）或兔白细胞介素-6（RaIL-6）融合并在细菌中表达。将 100μg 的 P2-gD/gC/gB-BoIL-6、P2-gD/gC/gB-RaIL-6、P2-gD/gC/gB、P2-gD/gC/gB 加 BoIL-6、P2-（gD-a）3-BoIL-6 或 P2-（gD-a）3 用 ISA 206 佐剂三次免疫肌肉注射，间隔 3 周。与 P2-gD/gC/gB 加 BoIL-6、P2-gD/gC/gB-RaIL-6 或其他制剂相比，P2-gD/gC/gB-BoIL-6 产生了更高滴度的 BoHV-1 特异性抗体、中和抗体、干扰素（IFN）-γ 和 IL-4。与灭活牛病毒性腹泻（BVD）-传染性牛鼻气管炎（IBR）疫苗相比，P2-gD/gC/gB-BoIL-6 诱导的抗体水平相似，但 IFN-γ 和 IL-4 的滴度显著更高。与 P2-gD/gC/gB 加 BoIL-6 或 P2-gD/gC/gB-RaIL-6 相比，接种 P2-gD/gC/gB-BoIL-6 的兔子在病毒攻击和再激活后肺部和气管中的病毒脱落和组织病变明显减少。P2-gD/gC/gB-BoIL-6 提供了与灭活疫苗相似的抗病毒脱落和组织发病的保护作用。该数据证实了多表位重组蛋白的安全性和免疫原性，是一种控制该疾病的潜在候选疫苗，特别是对怀孕牛。
