tetano
Editor, Senior Moderator
Hum Vaccin Immunother. 2015 Jun 19:0. [Epub ahead of print]
[h=1]Enhanced production of human influenza virus in PBS-12SF cells with a reduced interferon response.[/h] Carvajal-Yepes M[SUP]1[/SUP], Sporer KR, Carter JL, Colvin CJ, Coussens PM.
[h=3]Author information[/h]
[h=3]Abstract[/h] Influenza is one of the most important infectious diseases in humans. The best way to prevent severe illness caused by influenza infection is vaccination. Cell culture-derived influenza vaccines are being considered in addition to the widely used egg-based system in order to support the increasing seasonal demand and to be prepared in case of a pandemic. Cell culture based systems offer increased safety, capacity, and flexibility with reduced downstream processing relative to embryonated eggs. We have previously reported a chick embryo cell line, termed PBS-12SF, that supports replication of human and avian influenza A viruses to high titers (>10 [SUP]7[/SUP] PFU/ml) without the need for exogenous proteases or serum proteins. Viral infections in cells are limited by the Interferon (IFN) response typified by production of type I IFNs that bind to the IFNα/β receptor and activate an antiviral state. In this study, we investigated how neutralizing the interferon (IFN) response in PBS-12SF cells, via shRNA-mediated knock-down of IFNAR1 mRNA expression, affects influenza virus production. We were successful in knocking down ?90% of IFNAR1 protein expression by this method, resulting in a significant decrease in the response to recombinant chIFNα stimulation in PBS-12SF cells as shown by a reduction in expression of interferon-responsive genes when compared to control cells. Additionally; IFNAR1-knock-down cells displayed enhanced viral HA production and released more virus into cell culture supernatants than parental PBS-12SF cells.
[h=4]KEYWORDS:[/h] H1N1; Influenza; Interferon; Vaccine production; shRNA
PMID: 26090991 [PubMed - as supplied by publisher]
[h=1]Enhanced production of human influenza virus in PBS-12SF cells with a reduced interferon response.[/h] Carvajal-Yepes M[SUP]1[/SUP], Sporer KR, Carter JL, Colvin CJ, Coussens PM.
[h=3]Author information[/h]
[h=3]Abstract[/h] Influenza is one of the most important infectious diseases in humans. The best way to prevent severe illness caused by influenza infection is vaccination. Cell culture-derived influenza vaccines are being considered in addition to the widely used egg-based system in order to support the increasing seasonal demand and to be prepared in case of a pandemic. Cell culture based systems offer increased safety, capacity, and flexibility with reduced downstream processing relative to embryonated eggs. We have previously reported a chick embryo cell line, termed PBS-12SF, that supports replication of human and avian influenza A viruses to high titers (>10 [SUP]7[/SUP] PFU/ml) without the need for exogenous proteases or serum proteins. Viral infections in cells are limited by the Interferon (IFN) response typified by production of type I IFNs that bind to the IFNα/β receptor and activate an antiviral state. In this study, we investigated how neutralizing the interferon (IFN) response in PBS-12SF cells, via shRNA-mediated knock-down of IFNAR1 mRNA expression, affects influenza virus production. We were successful in knocking down ?90% of IFNAR1 protein expression by this method, resulting in a significant decrease in the response to recombinant chIFNα stimulation in PBS-12SF cells as shown by a reduction in expression of interferon-responsive genes when compared to control cells. Additionally; IFNAR1-knock-down cells displayed enhanced viral HA production and released more virus into cell culture supernatants than parental PBS-12SF cells.
[h=4]KEYWORDS:[/h] H1N1; Influenza; Interferon; Vaccine production; shRNA
PMID: 26090991 [PubMed - as supplied by publisher]