Department of Virology, Imperial College London, St. Mary's Campus, London, United Kingdom.
Vaccine. 2010 Apr 9;28(17):2973-9. doi: 10.1016/j.vaccine.2010.02.012. Epub 2010 Feb 25.
Most influenza vaccines are produced in chicken eggs but recent human influenza strains often do not grow well in this substrate. The PER.C6 cell line is an alternative platform for vaccine production. Here we demonstrate that PER.C6 cells faithfully propagate recent clinical isolates, without selecting for mutations in the HA gene. PER.C6 cells support the rescue of recombinant influenza viruses from cDNA. We used sequence data from a surveillance programme to generate a PR8-based seed virus with the HA and NA of a contemporary circulating H3N2 human strain, A/England/611/07 (E611) that did not itself grow in eggs. We engineered mutations that affected receptor-binding, G186V or L194P, into the E611 HA gene. Whilst the L194P mutation conferred efficient growth in eggs, G186V did not. The L194P mutation was also spontaneously selected during egg propagation of E611/PR8 7:1 recombinant virus. This suggests generation of a single recombinant vaccine seed might satisfy manufacturers that utilize either eggs or cells for vaccine production.
大多数流感疫苗是在鸡蛋中生产的，但最近的人类流感株在这种基质中往往生长不佳。PER.C6 细胞系是疫苗生产的替代平台。在这里，我们证明 PER.C6 细胞忠实地复制了最近的临床分离株，而不会选择在 HA 基因中发生突变。PER.C6 细胞支持从 cDNA 中拯救重组流感病毒。我们使用监测计划的序列数据，从 PR8 生成了一个基于种子病毒的种子病毒，其 HA 和 NA 来自当代流行的 H3N2 人类株 A/England/611/07（E611），该病毒本身不能在鸡蛋中生长。我们对 E611 HA 基因中的受体结合产生影响的突变，G186V 或 L194P 进行了工程改造。虽然 L194P 突变赋予了在鸡蛋中高效生长的能力，但 G186V 没有。在 E611/PR8 7:1 重组病毒的鸡蛋繁殖过程中，L194P 突变也自发选择。这表明生成单个重组疫苗种子可能会满足那些利用鸡蛋或细胞生产疫苗的制造商的要求。
