College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, China; Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, 510642, China; Guangdong Technological Engineering Research Center for Pet, Guangzhou, 510642, China.
College of Animal Science & Technology, Zhongkai University of Agriculture and Engineering, Guangzhou, 510225, China.
Vet Microbiol. 2021 Mar;254:108997. doi: 10.1016/j.vetmic.2021.108997. Epub 2021 Jan 27.
Canine influenza (CI) is a contagious respiratory disease in dogs, which poses a threat to canine health. A safe, high-yield vaccine seed virus is critical for CI vaccine development. We developed a PR8-based reassortant H3N2 canine influenza virus (RT CIV) using the reverse genetic method and evaluated its yield in canine kidney epithelial (MDCK) cells, Vero cells, and specific pathogen-free (SPF) chicken embryos. Mice and dogs were infected with RT CIV, and the pathogenicity was evaluated. The viral titers of RT CIV increased in MDCK cells, Vero cells, and SPF chicken embryos; the HA yield in SPF chicken embryos increased 4-fold. However, RT CIV was not lethal to mice, and it showed similar virulence as wild-type CIV. RT CIV also showed minimal pathogenicity in dogs, which manifested as mild fever and rhinorrhea for the first two days post-infection. Thus, RT CIV carrying the internal gene cassette from PR8 showed almost no pathogenicity in dogs. And the reassortant virus inactivated vaccine could provide complete protection against H3N2 CIV. To our knowledge, this is the first report on the pathogenicity of PR8-based reassortant H3N2 CIV in dogs. These studies are relevant for developing a high-yield and safe CI vaccine.
犬流感(CI)是一种犬类传染性呼吸道疾病,对犬类健康构成威胁。安全、高产量的疫苗种子病毒是犬流感疫苗开发的关键。我们使用反向遗传学方法开发了一种基于 PR8 的重配 H3N2 犬流感病毒(RT CIV),并评估了其在犬肾上皮(MDCK)细胞、Vero 细胞和无特定病原体(SPF)鸡胚中的产量。用 RT CIV 感染小鼠和犬,并评估其致病性。在 MDCK 细胞、Vero 细胞和 SPF 鸡胚中,RT CIV 的病毒滴度增加;SPF 鸡胚中的 HA 产量增加了 4 倍。然而,RT CIV 对小鼠没有致命性,其毒力与野生型 CIV 相似。RT CIV 在犬中也表现出最小的致病性,感染后前两天表现为轻度发热和流鼻涕。因此,携带 PR8 内部基因盒的 RT CIV 在犬中几乎没有致病性。重组病毒灭活疫苗可提供针对 H3N2 CIV 的完全保护。据我们所知,这是首次报道 PR8 基重组 H3N2 CIV 在犬中的致病性。这些研究对于开发高产、安全的 CI 疫苗具有重要意义。