• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文会议>Vaccine technology VI >INFLUENZA A VIRUS PROPAGATION IN MDCK: INTRACELLULAR VIRUS REPLICATION, VIRUS RELEASE AND CELL-CYCLE PREFERENTIAL INFECTION ANALYSIS
【24h】

INFLUENZA A VIRUS PROPAGATION IN MDCK: INTRACELLULAR VIRUS REPLICATION, VIRUS RELEASE AND CELL-CYCLE PREFERENTIAL INFECTION ANALYSIS

机译：在MDCK中感染病毒传播：细胞内病毒复制，病毒释放和细胞周期优先感染分析

获取原文
获取原文并翻译 | 示例
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Cell culture-based processes for vaccine manufacturing offer advantages over egg-based processes in terms of product uniformity and sterility, production time and scaling up capacity12. Regarding influenza vaccines, MDCK cells are one of the host cell lines currently used to manufacture licensed products; however, virus titers remain lower compared to those obtained in eggs and further increase of specific and volumetric yields is required. To identify bottlenecks in influenza A virus (IAV) production, we thoroughly studied IAV replication in MDCK cells. For this, we analyzed different features of the infection process such as viral RNA replication, intracellular localization of viral components, virus release and morphology of the particles, and the preferential infection in different cell-cycle phases. Using synchronous infections, we found that production of infectious particles dropped much earlier than the production of total particles. Furthermore, we found that the maximum virus release rate was reached when all viral RNA species attained their maximum intracellular concentration. Using qPCR we determined that the vRNA maximum concentration per cell was 10-fold higher than the specific viral titers obtained, indicating that vRNA concentration does not limit IAV particle assembly. When we evaluated the morphology of particles released using electron microscopy, we observed that a higher fraction of the viral particles produced at late times possess an abnormal morphology, concurring with the increased production of non-infectious viruses. Using imaging flow cytometry, we determined that the export of influenza viral genome segments (ribonucleoprotein complexes, vRNPs) from the nucleus to the cytoplasm strongly correlated with the onset of virus release. However, our results also suggest that the induction of apoptosis caused that virus assembly became deficient producing more non-infectious particles at late infection times. Lastly, using low MOI infections and imaging flow cytometry, we found that -in contrast to previous publications-IAV did not preferentially infect a specific cell cycle phase and no cell cycle arrest induction was observed during the time frame of the experiment (9 hpi). In summary, the data presented here offers a comprehensive overview of the dynamics of IAV infection in MDCK and might contribute to the development of molecular or cell culture-based strategies to improve IAV production in MDCK cells.
机译：在疫苗均匀性，无菌性，生产时间和扩大生产能力方面，基于细胞培养的疫苗生产工艺比蛋基工艺更具优势12。关于流感疫苗，MDCK细胞是目前用于制造许可产品的宿主细胞系之一。但是，病毒滴度仍比鸡蛋中的滴度低，需要进一步提高比容和体积产量。为了确定甲型流感病毒（IAV）产生的瓶颈，我们彻底研究了IAV在MDCK细胞中的复制。为此，我们分析了感染过程的不同特征，例如病毒RNA复制，病毒成分在细胞内的定位，病毒的释放和颗粒的形态以及在不同细胞周期阶段的优先感染。使用同步感染，我们发现传染性颗粒的产生比总颗粒的产生要早得多。此外，我们发现，当所有病毒RNA种类均达到其最大细胞内浓度时，病毒释放速率达到了最大值。使用qPCR，我们确定每个细胞的vRNA最大浓度比所获得的特定病毒滴度高10倍，表明vRNA浓度不限制IAV颗粒的组装。当我们评估使用电子显微镜释放的颗粒的形态时，我们观察到后期产生的更高比例的病毒颗粒具有异常的形态，这与非感染性病毒产量的增加有关。使用成像流式细胞仪，我们确定流感病毒基因组片段（核糖蛋白复合物，vRNPs）从细胞核到细胞质的出口与病毒释放的开始密切相关。但是，我们的结果还表明，凋亡的诱导导致病毒装配不足，在后期感染时产生更多的非感染性颗粒。最后，使用低MOI感染和成像流式细胞仪，我们发现-与以前的出版物相比，IAV不会优先感染特定的细胞周期阶段，并且在实验期间（9 hpi）未观察到细胞周期停滞诱导。总之，此处提供的数据全面概述了MDCK中IAV感染的动力学，可能有助于开发基于分子或细胞培养的策略以改善MDCK细胞中IAV的产生。

著录项

  • 来源
    《Vaccine technology VI》|2016年|210-210|共1页
  • 会议地点 Albufeira(PT)
  • 作者

    Gallo-Ramirez; Frensing T; Kupke S Bachmann;

  • 作者单位

    L.E, Max Planck Institute for Dynamics of Complex Technical Systems, Germany;

    Reichl, U , Max Planck Institute for Dynamics of Complex Technical Systems,Otto-von-Guericke-Universitaet, Germany;

    M, Fritzsche, S, Max Planck Institute for Dynamics of Complex Technical Systems, Germany;

  • 会议组织
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Influenza A virus; MDCK cells; replication dynamics;

    机译：甲型流感病毒； MDCK细胞；复制动态;

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号