English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Trypsin promotes efficient influenza vaccine production in MDCK cells by interfering with the antiviral host response

Seitz, C., Isken, B., Heynisch, B., Rettkowski, M., Frensing, T., & Reichl, U. (2012). Trypsin promotes efficient influenza vaccine production in MDCK cells by interfering with the antiviral host response. Applied Microbiology and Biotechnology, 93(2), 601-611. doi:10.1007/s00253-011-3569-8.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0013-8992-8 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-A3CE-1
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Seitz, C.1, Author              
Isken, B.1, Author              
Heynisch, B.1, Author              
Rettkowski, M.1, Author              
Frensing, T.1, Author              
Reichl, U.1, 2, Author              
Affiliations:
1Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society, ou_1738140              
2Otto-von-Guericke-Universität Magdeburg, ou_1738156              

Content

show
hide
Free keywords: Influenza, vaccine production, MDCK cells, trypsin, interferon
 Abstract: Trypsin is commonly used in Madin-Darby canine kidney (MDCK) cell culture-based influenza vaccine production to facilitate virus infection by proteolytic activation of viral haemagglutinin, which enables multi-cycle replication. In this study, we were able to demonstrate that trypsin also interferes with pathogen defence mechanisms of host cells. In particular, a trypsin concentration of 5 BAEE U/mL (4.5 µg/ml porcine trypsin) used in vaccine manufacturing strongly inhibited interferon (IFN) signalling by proteolytic degradation of secreted IFN. Consequently, absence of trypsin during infection resulted in a considerably stronger induction of IFN signalling and apoptosis, which significantly reduced virus yields. Under this condition, multi-cycle virus replication in MDCK cells was not prevented but clearly delayed. Therefore, incomplete infection can be ruled out as the reason for the lower virus titres. However, suppression of IFN signalling by overexpression of viral IFN antagonists (influenza virus PR8-NS1, rabies virus phosphoprotein) partially rescued virus titres in the absence of trypsin. In addition, virus yields could be almost restored by using the influenza strain A/WSN/33 in combination with fetal calf serum (FCS). For this strain FCS enabled trypsin-independent fast propagation of virus infection, probably outrunning cellular defence mechanisms and apoptosis induction in the absence of trypsin. Overall, addition of trypsin provided optimal conditions for high yield vaccine production in MDCK cells by two means. On the one hand, proteolytic degradation of IFN keeps cellular defence at a low level. On the other hand, enhanced virus spreading enables viruses to replicate before the cellular response becomes fully activated. copyright Springer-Verlag 2011 [accessed November 30th 2011]

Details

show
hide
Language(s): eng - English
 Dates: 2012
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 573862
DOI: 10.1007/s00253-011-3569-8
Other: 4/12
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Applied Microbiology and Biotechnology
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Heidelberg : Springer-Verlag
Pages: - Volume / Issue: 93 (2) Sequence Number: - Start / End Page: 601 - 611 Identifier: ISSN: 0175-7598
CoNE: /journals/resource/954928543201