Department of Infection Biology, Faculty of Health and Life Sciences, University of Liverpool, Leahurst Campus, Neston, Cheshire CH64 7TE, United Kingdom.
Vaccine. 2011 Nov 28;29(51):9493-8. doi: 10.1016/j.vaccine.2011.10.024. Epub 2011 Oct 19.
Throughout the world, avian metapneumovirus (AMPV) infection of subtype A is principally controlled by two live vaccines both derived from UK field strain #8544. Improvements of those vaccines by use of reverse genetics technology was found to be hampered by the inability of #8544 to replicate in the commonly exploited Vero cell based reverse genetics system. A systematic reverse genetics based genome modification of a DNA copy of #8544, employing sequence data from a Vero grown, #8544 derived, live vaccine; was used to determine mutations required to facilitate virus recovery and replication in Vero cells. This identified a single coding substitution in the M2:2 reading frame as responsible. Furthermore, ablation of M2:2 was found to elicit the same outcome. M2:2 sequence analysis of seven AMPVs found Vero cell adaption to be associated with non similar amino acid changes in M2:2. The study shows that M2:2 modification of field virus #8544 will enable research leading to improved vaccines. This may have more general application to other AMPV field strains.
在全球范围内，主要由两种源自英国田间分离株#8544 的活疫苗来控制禽偏肺病毒（AMPV）亚型 A 的感染。通过反向遗传技术对这些疫苗进行改进，发现由于#8544 无法在常用的基于 Vero 细胞的反向遗传系统中复制，因此受到阻碍。对#8544 的 DNA 拷贝进行了基于反向遗传学的系统基因组修饰，使用了从在 Vero 细胞中生长的、源自#8544 的活疫苗获得的序列数据，以确定促进病毒在 Vero 细胞中恢复和复制所需的突变。这确定了 M2:2 阅读框中的单个编码取代是负责的。此外，还发现缺失 M2:2 会产生相同的结果。对在 Vero 细胞中适应的七种 AMPV 的 M2:2 序列分析表明，与 M2:2 中的非相似氨基酸变化相关。该研究表明，对田间病毒#8544 的 M2:2 修饰将能够进行研究，从而开发出更好的疫苗。这可能更普遍适用于其他 AMPV 田间株。
