Ezrin and HNRNP expression correlate with increased virus release rate and early onset of virus‐induced apoptosis of MDCK suspension cells

Abstract

With the aim to adapt high-yield adherent cell lines to suspension growth, Madin Darby canine kidney (MDCK) suspension cells were developed recently that achieved comparable influenza virus yields despite an early induction of apoptosis compared to the parental adherent cell line. For both cell lines, a comprehensive study under comparable infection conditions is performed comprising information on: time course of viral infection, antiviral state of cells, virus-induced apoptosis, and virus-induced cellular protein expression for early and late infection with influenza A/PuertoRico/8/34 H1N1. The proteomic analysis is performed with 2D differential gel electrophoreses followed by mass spectrometry. Based on flow cytometric data and on the differential expression of various stress and apoptosis-related proteins, the earlier onset of virus-induced apoptosis is confirmed for suspension cells. Surprisingly, the data indicated an increased virus release rate for suspension cells. These observations correlate with an increased expression of the apical marker protein ezrin, known to play a role in influenza-induced cytoskeletal rearrangement, and the differential expression of heterogeneous nuclear ribonucleoproteins, known to bind actively influenza viral proteins and play a central role in regulating gene expression. Based on these findings, additional studies towards the design of MDCK suspension cells with further increase in influenza virus yields will be performed.

Inhibition of H1N1 influenza virus by selenium nanoparticles loaded with zanamivir through p38 and JNK signaling pathways

“…It has been reported that H1N1 inuenza virus induces apoptosis of cells and caspase-3 participates in the process. 47,48 Caspase-3 plays as a pivotal role in apoptosis, which can be activated during virus infection. It is responsible for the proteolytic cleavage of many critical proteins, such as the poly-ADP-ribose polymerase (PARP).…”

Reduction in Interferon-Stimulated Genes Contributes to High-Yield Production of Influenza Virus in Suspension MDCK Cells

“…MDCK suspension cells obtained by serum-free adaption culture have a faster growth rate, allowing for mass production, which has been reported to produce influenza vaccines [ 12 , 13 , 14 , 15 , 16 , 17 ]. However, most research focuses on cell growth and metabolic regulation, and limited studies have examined the mechanism of cell immune response during virus production [ 16 , 18 ].…”