Susta Leonardo, He Ying, Hutcheson Jessica M, Lu Yangqing, West Franklin D, Stice Steven L, Yu Ping, Abdo Zaid, Afonso Claudio L
US National Poultry Research Center, Exotic and Emerging Avian Viral Diseases Research Unit, Southeast Poultry Research Laboratory, Athens, GA, 30605, USA.
Present address: Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, ON, N1G 2 W1, Canada.
Virol J. 2016 Dec 5;13(1):205. doi: 10.1186/s12985-016-0659-3.
Newcastle disease (ND), caused by Newcastle disease virus (NDV), is a devastating disease of poultry and wild birds. ND is prevented by rigorous biocontainment and vaccination. One potential approach to prevent spread of the virus is production of birds that show innate resistance to NDV-caused disease. Induced pluripotent stem cell (iPSC) technology allows adult cells to be reprogrammed into an embryonic stem cell-like state capable of contributing to live offspring and passing on unique traits in a number of species. Recently, iPSC approaches have been successfully applied to avian cells. If chicken induced pluripotent stem cells (ciPSCs) are genetically or epigenetically modified to resist NDV infection, it may be possible to generate ND resistant poultry. There is limited information on the potential of ciPSCs to be infected by NDV, or the capacity of these cells to become resistant to infection. The aim of the present work was to assess the characteristics of the interaction between NDV and ciPSCs, and to develop a selection method that would increase tolerance of these cells to NDV-induced cellular damage.
Results showed that ciPSCs were permissive to infection with NDV, and susceptible to virus-mediated cell death. Since ciPSCs that survived infection demonstrated the ability to recover quickly, we devised a system to select surviving cells through multiple infection rounds with NDV. ciPSCs that sustained 9 consecutive infections had a statistically significant increase in survival (up to 36 times) compared to never-infected ciPSCs upon NDV infection (tolerant cells). Increased survival was not caused by a loss of permissiveness to NDV replication. RNA sequencing followed by enrichment pathway analysis showed that numerous metabolic pathways where differentially regulated between tolerant and never-infected ciPSCs.
Results demonstrate that ciPSCs are permissive to NDV infection and become increasingly tolerant to NDV under selective pressure, indicating that this system could be applied to study mechanisms of cellular tolerance to NDV.
新城疫（ND）由新城疫病毒（NDV）引起，是一种对家禽和野生鸟类具有毁灭性的疾病。通过严格的生物防护和疫苗接种来预防新城疫。一种预防病毒传播的潜在方法是培育对新城疫病毒引起的疾病具有先天抵抗力的禽类。诱导多能干细胞（iPSC）技术可使成体细胞重编程为类似胚胎干细胞的状态，能够产生存活后代并在多个物种中传递独特性状。最近，iPSC方法已成功应用于禽类细胞。如果对鸡诱导多能干细胞（ciPSC）进行基因或表观遗传修饰以抵抗新城疫病毒感染，那么有可能培育出抗新城疫的家禽。关于ciPSC被新城疫病毒感染的可能性或这些细胞对感染产生抗性的能力的信息有限。本研究的目的是评估新城疫病毒与ciPSC之间相互作用的特征，并开发一种选择方法，以提高这些细胞对新城疫病毒诱导的细胞损伤的耐受性。
结果表明，ciPSC对新城疫病毒感染敏感，且易受病毒介导的细胞死亡影响。由于感染后存活的ciPSC显示出快速恢复的能力，我们设计了一个系统，通过用新城疫病毒进行多次感染来选择存活细胞。与未感染的ciPSC相比，连续9次感染的ciPSC在新城疫病毒感染后（耐受细胞）存活率有统计学显著提高（高达36倍）。存活率的提高不是由于对新城疫病毒复制的易感性丧失。RNA测序及随后的富集通路分析表明，耐受和未感染的ciPSC之间有许多代谢通路存在差异调节。
结果表明，ciPSC对新城疫病毒感染敏感，且在选择压力下对新城疫病毒的耐受性越来越高，这表明该系统可用于研究细胞对新城疫病毒的耐受机制。
