College of Animal Science and Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an, 271018, China.
Institute of Poultry Science, Shandong Academy of Agricultural Science, Jinan, 250023, China.
Antiviral Res. 2017 Nov;147:1-10. doi: 10.1016/j.antiviral.2017.04.019. Epub 2017 Apr 29.
The present study is focused on investigating the immunoprotective effects of CpG-ODN/Poly(I:C) combined with the viral glycoprotein gp90 protein against reticuloendotheliosis virus (REV) infection in chickens. REV's gp90 gene was amplified from the REV-infected cells and expressed in Escherichia coli (E.coli). The expressed products, upon purification, were inoculated into 7-day-old chickens with PBS, CpG-ODN or Poly(I:C) adjuvant; Two booster inoculations were then conducted, and then each chicken was challenged. The presence of REV-antibodies in serum was determined weekly after the first vaccination. The viremia and immunosuppressive effects of REV infection were also monitored after the challenge. The neutralizing effects of the antisera were tested in vitro. The results showed that the recombinant gene containing REV gp90 gene was expressed into the recombinant protein with a size of 51 Kilo Dalton (KD), which could be recognized by a monoclonal antibody (MAb) against the gp90 protein. The viremia and immunosuppressive effects of avian influenza virus (AIV) vaccine caused by REV challenge in CpG-ODN group and in Poly(I:C) group were dramatically decreased. REV antibody with low titers was induced in gp90 group and the inoculated chickens were partly protected. Compared with those in gp90 group, the titers and the positive ratios of REV antibody in CpG+gp90 group were significantly increased, whereas the viremia and immunosuppressive effects of AIV vaccine caused by REV infection were significantly decreased. In the Poly(I:C) +gp90 group, the viremia and immunosuppressive effects caused by REV infection were also dramatically decreased, although REV antibody responses were softly increased. The diluted antisera from the vaccinated chickens in both groups could completely inhibit the replication of REV in chick fibroblast cells (CEF). Hence, it can be concluded that CpG-ODN or the Poly(I:C) adjuvant can enhance the antiviral effects of the REV subunit vaccine against REV infection, which may result from different mechanisms.
本研究旨在探讨 CpG-ODN/Poly(I:C)与病毒糖蛋白 gp90 蛋白联合用于预防鸡传染性网状内皮组织增生病病毒 (REV)感染的免疫保护作用。从感染 REV 的细胞中扩增 REV 的 gp90 基因,并在大肠杆菌 (E.coli) 中表达。表达产物经纯化后,用 PBS、CpG-ODN 或 Poly(I:C)佐剂接种 7 日龄鸡;然后进行两次加强接种,然后每只鸡进行攻毒。第一次接种后每周检测血清中 REV 抗体的存在。攻毒后还监测了 REV 感染的病毒血症和免疫抑制作用。体外测试了抗血清的中和作用。结果表明,含有 REV gp90 基因的重组基因表达为大小为 51 千道尔顿 (KD) 的重组蛋白,可被针对 gp90 蛋白的单克隆抗体 (MAb) 识别。CpG-ODN 组和 Poly(I:C) 组 REV 攻毒引起的禽流感病毒 (AIV) 疫苗的病毒血症和免疫抑制作用明显降低。在 gp90 组中诱导出低滴度的 REV 抗体,接种鸡得到部分保护。与 gp90 组相比,CpG+gp90 组的 REV 抗体滴度和阳性率明显升高,而 REV 感染引起的 AIV 疫苗的病毒血症和免疫抑制作用明显降低。在 Poly(I:C)+gp90 组中,REV 感染引起的病毒血症和免疫抑制作用也明显降低,尽管 REV 抗体反应略有增加。两组接种鸡的稀释抗血清均可完全抑制 REV 在鸡成纤维细胞 (CEF) 中的复制。因此,可以得出结论,CpG-ODN 或 Poly(I:C) 佐剂可以增强 REV 亚单位疫苗对 REV 感染的抗病毒作用,这可能是由于不同的机制。