- 作者: Tsai-Teng Tzeng, Po-Ling Chen, Tsai-Chuan Weng, Shin-Yi Tsai, Chia-Chun Lai, Hsin-I Chou, Pin-Wen Chen, Chia-Chun Lu, Ming-Tsan Liu, Wang-Chou Sung, Min-Shi Lee and Alan Yung-Chih Hu
- 作者服務機構: 1. National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), 35 Keyan Road, Zhunan, Miaoli County, 35053, Taiwan 2. Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu, Taiwan 3. College of Life Science, National Tsing Hua University, Hsinchu, Taiwan 4. Centers for Disease Control, Ministry of Health and Welfare, Taipei, 689, Taiwan
- 中文摘要:
- 英文摘要:
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. - 中文關鍵字:
- 英文關鍵字: H7N9, Highly pathogenic avian influenza, Candidate vaccine virus, Suspension MDCK cells, Chemically defined medium, Reverse genetics