Application of an Inclined Settler for Cell Culture-Based Influenza A Virus Production in Perfusion Mode

influenza virus have been successfully deployed using a variety of animal cell lines in batch, fed-batch and perfusion culture. For suspension cells, most studies reported on membrane-based cell retention device usually causes the accumulation of the virus in a bioreactor in perfusion mode.

Virus harvest continuously aims to improve productivity, a settler tend evaluated for influenza A virus (IAV) production using the birds AGE1.CR.pIX suspension cell lines. settlers tend to provide a lot of advantages because they are scalable, robust, and comply with cGMP regulations, for example, for the manufacture of recombinant proteins. perfusion levels up to 3000 L / day has been reported. In our study, the successful growth of cells AGE1.CR.pIX to 50 × 106 cells / mL and a cell retention efficiency exceeding 96% was obtained with the settlers cooled to room temperature.

No virus observed retention. A total of 5.4 to 6.5 × 1013 virions produced while control experiments with ATF system equaled 1.9 × 1013 virions. For infections of the 25 × 106 cells / mL, virus-specific cells produces up to 3474 virions / cell was obtained, approximately 5-fold higher than for cultivation based ATF conducted as controls (723 virions / cell). Trypsin activity proved to have a major impact on the dynamics of cell growth after a subsequent infection cell retention device, especially on a cell concentration of 50 × 106 cells / mL.

Further control experiments carried out by settlers acoustic show that virus production increased with the heat exchanger of the settlers tend to be operated at 27 ° C. In short, cell culture-based virus production in perfusion mode with sloping settlers and continuous harvest can drastically improve results and may IAV results of other viruses. To our knowledge, this is the first report that shows the potential of this device for the production of viral vaccines.

Application of an Inclined Settler for Cell Culture-Based Influenza A Virus Production in Perfusion Mode
Application of an Inclined Settler for Cell Culture-Based Influenza A Virus Production in Perfusion Mode

avoidance mechanisms of type I interferon response by influenza A virus

Type I interferon (IFN) represent the first line of host defense against influenza virus infection, and precise control of type I IFN response is the central event of immune defense against influenza virus infection. Influenza virus is one of the main causes of respiratory tract infections in humans and are responsible for seasonal epidemics and pandemics occasionally, leading to a serious threat to global human health due to antigenic variation and interspecies transmission of them.

Although the host cells have evolved mechanisms of antiviral advanced by sensing products of influenza virus and trigger signaling cascades resulting in the secretion of IFN type I (IFN-α / β), influenza viruses have developed strategies to counter these mechanisms and avoid the type I IFNs in response, for example, with induces host shut-off, or by arranging polyubiquitination viral proteins and host. This review will summarize the current knowledge about how the host cells recognize influenza virus to induce type I IFN response and the strategies used to prevent influenza virus type I IFN signaling pathways, which will help to antiviral and vaccine development.

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In the last months of 2019, an outbreak of fatal respiratory disease that began in Wuhan, China, and quickly spread to other parts of the world. It named COVID-19, and to date, thousands of cases of infection and deaths reported worldwide. The disease is associated with a variety of symptoms that make it difficult to diagnose accurately.

Previous SARS pandemic in 2003, researchers found that patients with fever, cough, or sore throat has influenza virus-positive rate of 5%. These findings triggered in our minds that the various symptoms and also the relatively high prevalence of death in our patients may be due to co-infection with other viruses.