Zegpi R A, Breedlove C, van Santen V L, Rasmussen-Ivey C R, Toro H
A Department of Pathobiology, Auburn University College of Veterinary Medicine, Auburn, AL 36849.
B Department of Biological Sciences, Auburn University College of Sciences and Mathematics, Auburn, AL 36849.
Avian Dis. 2017 Jun;61(2):221-228. doi: 10.1637/11537-111416-Reg.1.
We previously demonstrated that adaptation of an embryo-attenuated infectious bronchitis virus (IBV) Arkansas (Ark) Delmarva Poultry Industry (DPI)-derived vaccine to chicken embryo kidney (CEK) cells (CEKp7) shifted the virus population towards homogeneity in spike (S) and nonstructural protein genes. Moreover, the typical Ark vaccine subpopulations emerging in chickens vaccinated with commercial Ark vaccines were not detected in chickens vaccinated with CEKp7, indicating that kidney-cell adaptation drastically increased the stability of the vaccine virus population in chickens. In the current study both conventional and next-generation sequencing results show that the changes achieved during CEK adaptation remained after five back passages in embryonated chicken egg (ECE). In a first protection study 1-day-old chickens were vaccinated with 10 or 10 50% embryo infectious doses (EID)/chicken of the second ECE back passage of CEKp7 (CEKp7e2) and demonstrated protection against Ark virulent (10 EID) challenge. In a second protection trial, protection by CEKp7e2 was compared with protection conferred by an attenuated commercial ArkDPI-derived vaccine different from that which the CEK-adapted virus originated. All vaccinated chicken groups showed a significant reduction of respiratory signs and viral load after Ark virulent challenge compared to unvaccinated-challenged controls. In CEKp7e2 vaccinated chickens viral subpopulations different from the challenge virus were detected after challenge in a marginal number (7%-8%) of chickens. In contrast, IBV S1 sequences that differed from the predominant population in the challenge virus were detected after challenge in a large number (77%) of chickens vaccinated with the commercial Ark attenuated vaccine. The CEK-adapted IBV ArkDPI-derived vaccine is a stable and effective vaccine, which drastically reduces the emergence of Ark-like viruses both at vaccination and after challenge.
我们之前证明,一种源自胚胎减毒传染性支气管炎病毒(IBV)阿肯色州(Ark)德尔马瓦家禽业(DPI)的疫苗适应鸡胚肾(CEK)细胞(CEKp7)后,病毒群体在刺突(S)和非结构蛋白基因方面趋向同质化。此外,在用商业Ark疫苗接种的鸡中出现的典型Ark疫苗亚群,在用CEKp7接种的鸡中未检测到,这表明肾细胞适应极大地提高了疫苗病毒群体在鸡体内的稳定性。在当前研究中,传统测序和下一代测序结果均显示,CEK适应过程中实现的变化在鸡胚(ECE)中连续传代五次后依然存在。在第一项保护研究中,1日龄雏鸡接种了第二代ECE传代的CEKp7(CEKp7e2),剂量为10或10个50%胚感染剂量(EID)/只鸡,结果显示对Ark强毒株(10个EID)攻击具有保护作用。在第二项保护试验中,将CEKp7e2的保护作用与一种源自ArkDPI的减毒商业疫苗的保护作用进行了比较,该商业疫苗与CEK适应病毒所源自的疫苗不同。与未接种疫苗并接受攻击的对照组相比,所有接种疫苗的鸡群在受到Ark强毒株攻击后,呼吸道症状和病毒载量均显著降低。在接种CEKp7e2的鸡中,攻击后在少数(7%-8%)鸡中检测到与攻击病毒不同的病毒亚群。相比之下,在接种商业Ark减毒疫苗的大量(77%)鸡中,攻击后检测到与攻击病毒中优势群体不同的IBV S1序列。适应CEK的源自IBV ArkDPI的疫苗是一种稳定有效的疫苗,在接种疫苗时以及攻击后都能大幅减少类Ark病毒的出现。