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悬浮 MDCK 细胞中高增长 H7N9 流感大流行前候选疫苗病毒的开发。

Development of high-growth influenza H7N9 prepandemic candidate vaccine viruses in suspension MDCK cells.

机构信息

National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), 35 Keyan Road, Zhunan, Miaoli County, 35053, Taiwan.

Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu, Taiwan.

出版信息

J Biomed Sci. 2020 Apr 2;27(1):47. doi: 10.1186/s12929-020-00645-y.

Abstract

BACKGROUND

Influenza vaccine manufacturers traditionally use egg-derived candidate vaccine viruses (CVVs) to produce high-yield influenza viruses for seasonal or pandemic vaccines; however, these egg-derived CVVs need an adaptation process for the virus to grow in mammalian cells. The low yields of cell-based manufacturing systems using egg-derived CVVs remain an unsolved issue. This study aimed to develop high-growth cell-derived CVVs for MDCK cell-based vaccine manufacturing platforms.

METHODS

Four H7N9 CVVs were generated in characterized Vero and adherent MDCK (aMDCK) cells. Furthermore, reassortant viruses were amplified in adherent MDCK (aMDCK) cells with certification, and their growth characteristics were detected in aMDCK cells and new suspension MDCK (sMDCK) cells. Finally, the plaque-forming ability, biosafety, and immunogenicity of H7N9 reassortant viruses were evaluated.

RESULTS

The HA titers of these CVVs produced in proprietary suspension MDCK (sMDCK) cells and chicken embryos were 2- to 8-fold higher than those in aMDCK cells. All H7N9 CVVs showed attenuated characteristics by trypsin-dependent plaque assay and chicken embryo lethality test. The alum-adjuvanted NHRI-RG5 (derived from the fifth wave H7N9 virus A/Guangdong/SP440/2017) vaccine had the highest immunogenicity and cross-reactivity among the four H7N9 CVVs. Finally, we found that AddaVax adjuvant improved the cross-reactivity of low pathogenic H7N9 virus against highly pathogenic H7N9 viruses.

CONCLUSIONS

Our study indicates that cell-derived H7N9 CVVs possessed high growth rate in new sMDCK cells and low pathogenicity in chicken embryo, and that CVVs generated by this platform are also suitable for both cell- and egg-based prepandemic vaccine production.

摘要

背景

流感疫苗制造商传统上使用源自鸡蛋的候选疫苗病毒(CVV)来生产用于季节性或大流行疫苗的高产量流感病毒;然而,这些源自鸡蛋的 CVV 需要适应过程才能在哺乳动物细胞中生长。使用源自鸡蛋的 CVV 的基于细胞的制造系统的低产量仍然是一个未解决的问题。本研究旨在开发用于 MDCK 细胞基疫苗制造平台的高增长细胞衍生 CVV。

方法

在有特征的 Vero 和贴壁 MDCK(aMDCK)细胞中生成了 4 种 H7N9 CVV。此外,在贴壁 MDCK(aMDCK)细胞中扩增重组病毒,并在 aMDCK 细胞和新悬浮 MDCK(sMDCK)细胞中检测其生长特性。最后,评估了 H7N9 重组病毒的蚀斑形成能力、生物安全性和免疫原性。

结果

在专有的悬浮 MDCK(sMDCK)细胞和鸡胚中产生的这些 CVV 的 HA 滴度比在 aMDCK 细胞中高 2-8 倍。所有 H7N9 CVV 通过依赖胰蛋白酶的蚀斑测定和鸡胚致死性试验显示出减毒特征。用铝佐剂配制的 NHRI-RG5(源自第五波 H7N9 病毒 A/广东/SP440/2017)疫苗在 4 种 H7N9 CVV 中具有最高的免疫原性和交叉反应性。最后,我们发现 AddaVax 佐剂提高了低致病性 H7N9 病毒对高致病性 H7N9 病毒的交叉反应性。

结论

我们的研究表明,源自细胞的 H7N9 CVV 在新的 sMDCK 细胞中具有高增长率和在鸡胚中的低致病性,并且该平台生成的 CVV 也适用于细胞和基于鸡蛋的大流行前疫苗生产。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b904/7115086/271d23f4c221/12929_2020_645_Fig1_HTML.jpg

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