MDCK cells that express proteases TMPRSS2 and HAT provide a cell system to propagate influenza viruses in the absence of trypsin and to study cleavage of HA and its inhibition

Vaccine ◽  
2009 ◽  
Vol 27 (45) ◽  
pp. 6324-6329 ◽  
Author(s):  
Eva Böttcher ◽  
Catharina Freuer ◽  
Torsten Steinmetzer ◽  
Hans-Dieter Klenk ◽  
Wolfgang Garten
Keyword(s):  
Mdck Cells ◽  
Cell System ◽  
Influenza Viruses ◽  
A Cell

Proteomic Characterization of Protein-Protein Interactions

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Veenstra TD ◽  
Keyword(s):  
Protein Interactions ◽  
Disease Diagnosis ◽  
Cell System ◽  
Protein Protein Interactions ◽  
Novel Technologies ◽  
Cell Functions ◽  
Single Protein ◽  
A Cell ◽  
Molecular Components

Identifying all the molecular components within a living cell is the first step into understanding how it functions. To further understand how a cell functions requires identifying the interactions that occur between these components. This fact is especially relevant for proteins. No protein within a human cell functions on its own without interacting with another biomolecule - usually another protein. While Protein-Protein Interactions (PPI) have historically been determined by examining a single protein per study, novel technologies developed over the past couple of decades are enabling high-throughput methods that aim to describe entire protein networks within cells. In this review, some of the technologies that have led to these developments are described along with applications of these techniques. Ultimately the goal of these technologies is to map out the entire circuitry of PPI within human cells to be able to predict the global consequences of perturbations to the cell system. This predictive capability will have major impacts on the future of both disease diagnosis and treatment.

Patterning of Lectins of Vero and MDCK Cells and Influenza Viruses

2005 ◽  
pp. 49-70 ◽  
Author(s):  
E Rapoport ◽  
N Bovin ◽  
L Mochalova ◽  
J Romanova ◽  
H Gabius
Keyword(s):  
Mdck Cells ◽  
Influenza Viruses

scholarly journals 2P209 A cell culture condition which prevents anoikis of MDCK cells

2004 ◽  
Vol 44 (supplement) ◽  
pp. S162
Author(s):  
Y. Emoto ◽  
C. Imafuku ◽  
K. Tawada
Keyword(s):  
Cell Culture ◽  
Culture Condition ◽  
Mdck Cells ◽  
A Cell ◽  
Cell Culture Condition

scholarly journals Overcoming the Challenges for a Mass Manufacturing Machine for the Assembly of PEMFC Stacks

Machines ◽  
2019 ◽  
Vol 7 (4) ◽  
pp. 66 ◽  
Author(s):  
Porstmann ◽  
Wannemacher ◽  
Richter
Keyword(s):  
Fuel Cell ◽  
Economies Of Scale ◽  
Polymer Electrolyte Membrane ◽  
Cell System ◽  
Active Components ◽  
Fuel Cell Stack ◽  
Electrolyte Membrane ◽  
Modular Concept ◽  
A Cell ◽  
Manufacturing Machine

One of the major obstacles standing in the way of a break-through in fuel cell technology is its relatively high costs compared to well established fossil-based technologies. The reasons for these high costs predominantly lie in the use of non-standardized components, complex system components, and non-automated production of fuel cells. This problem can be identified at multiple levels, for example, the electrochemically active components of the fuel cell stack, peripheral components of the fuel cell system, and eventually on the level of stack and system assembly. This article focused on the industrialization of polymer electrolyte membrane fuel cell (PEMFC) stack components and assembly. To achieve this, the first step is the formulation of the requirement specifications for the automated PEMFC stack production. The developed mass manufacturing machine (MMM) enables a reduction of the assembly time of a cell fuel cell stack to 15 minutes. Furthermore the targeted automation level is theoretically capable of producing up to 10,000 fuel cell stacks per year. This will result in a ~50% stack cost reduction through economies of scale and increased automation. The modular concept is scalable to meet increasing future demand which is essential for the market ramp-up and success of this technology.

scholarly journals Fusion and Infection of Influenza and Sendai Viruses as Modulated by Dextran Sulfate: A Comparative Study

2001 ◽  
Vol 21 (3) ◽  
pp. 293-304 ◽  
Author(s):  
João Ramalho-Santos ◽  
Maria C. Pedroso de Lima
Keyword(s):  
Molecular Weight ◽  
Dextran Sulfate ◽  
Sendai Virus ◽  
Mdck Cells ◽  
Inhibitory Effect ◽  
Low Ph ◽  
Influenza Viruses ◽  
Target Cells ◽  
Fusion Activity ◽  
Viral Envelopes

We have directly compared the effect of two types of dextran sulfate with distinct molecular weights (500 kDa and 5 kDa) on the fusion activity and infectivity of both Sendai and influenza viruses, two lipid-enveloped viruses that differ in their routes of entry into target cells. To correlate membrane merging and infectivity MDCK cells were used as targets for the viruses in both approaches. In either case pronounced inhibition of virus–cell interactions by dextran sulfate was only observed at low pH, even though Sendai virus fuses maximally at pH 7.4. Although membrane merging could not be fully abolished, the inhibitory effect was always greater when the higher molecular weight dextran sulfate was used. The presence of this residual fusion activity, that could not be reduced even with high concentrations of agent, suggests that a limited number of binding sites for dextran sulfate may exist on the viral envelopes. The compounds also inhibited fusion of bound virions, and all results could be reproduced using erythrocyte ghosts as target membranes in the fusion assay, instead of MDCK cells. In agreement with these observations only the infectivity of influenza virus (which requires a low pH-dependent step to enter target cells) was affected by dextran sulfate, again the higher molecular weight compound showing a more pronounced inhibitory effect.

Specific coupling of a cation-sensing receptor to G protein alpha-subunits in MDCK cells

1997 ◽  
Vol 273 (1) ◽  
pp. F129-F135 ◽  
Author(s):  
J. M. Arthur ◽  
G. P. Collinsworth ◽  
T. W. Gettys ◽  
L. D. Quarles ◽  
J. R. Raymond
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Plasma Membranes ◽  
Mdck Cells ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Cellular Activation ◽  
Protein Activation ◽  
A Cell ◽  
Cation Sensing

Extracellular cations such as Ca2+ stimulate a G protein-coupled, cation-sensing receptor (CaR). We used microphysiometry to determine whether an extracellular cation-sensing mechanism exists in Madin-Darby canine kidney (MDCK) cells. The CaR agonists Ca2+ and Gd3+ caused cellular activation in a concentration-dependent manner. mRNA for the CaR was identified by reverse transcription and polymerase chain reaction (PCR) using nested CaR-specific primers, identification of an appropriately located restriction site, and sequencing of the subcloned fragment obtained by PCR. G protein activation was evaluated using the GTP photoaffinity label [alpha-32P]GTP azidoanalide (AA-GTP). After stimulation with Gd3+ and cross-linking, plasma membranes were solubilized and immunoprecipitated with antisera specific for Gq/11 alpha and Gi alpha family members. Gd3+ increased incorporation of AA-GTP into Gq/11 alpha precipitates by 146 +/- 48% and into G alpha i-2 and G alpha i-3 to a lesser extent but not into G alpha i-1. Direct effects of Gd3+ on the G proteins were ruled out using partially purified mammalian G proteins expressed in Escherichia coli or Sf9 cells. We conclude that MDCK cells possess a cell-surface CaR that activates Gq/11 alpha, G alpha i-2, and G alpha i-3 but not G alpha i-1.

scholarly journals In Vitro Antiviral Activity of Favipiravir (T-705) against Drug-Resistant Influenza and 2009 A(H1N1) Viruses

2010 ◽  
Vol 54 (6) ◽  
pp. 2517-2524 ◽  
Author(s):  
Katrina Sleeman ◽  
Vasiliy P. Mishin ◽  
Varough M. Deyde ◽  
Yousuke Furuta ◽  
Alexander I. Klimov ◽  
...  
Keyword(s):  
Influenza A ◽  
Seasonal Influenza ◽  
Mdck Cells ◽  
Antiviral Effect ◽  
Plaque Formation ◽  
Influenza Viruses ◽  
New Drugs ◽  
Yield Reduction ◽  
Wide Range

ABSTRACT Favipiravir (T-705) has previously been shown to have a potent antiviral effect against influenza virus and some other RNA viruses in both cell culture and in animal models. Currently, favipiravir is undergoing clinical evaluation for the treatment of influenza A and B virus infections. In this study, favipiravir was evaluated in vitro for its ability to inhibit the replication of a representative panel of seasonal influenza viruses, the 2009 A(H1N1) strains, and animal viruses with pandemic (pdm) potential (swine triple reassortants, H2N2, H4N2, avian H7N2, and avian H5N1), including viruses which are resistant to the currently licensed anti-influenza drugs. All viruses were tested in a plaque reduction assay with MDCK cells, and a subset was also tested in both yield reduction and focus inhibition (FI) assays. For the majority of viruses tested, favipiravir significantly inhibited plaque formation at 3.2 μM (0.5 μg/ml) (50% effective concentrations [EC50s] of 0.19 to 22.48 μM and 0.03 to 3.53 μg/ml), and for all viruses, with the exception of a single dually resistant 2009 A(H1N1) virus, complete inhibition of plaque formation was seen at 3.2 μM (0.5 μg/ml). Due to the 2009 pandemic and increased drug resistance in circulating seasonal influenza viruses, there is an urgent need for new drugs which target influenza. This study demonstrates that favipiravir inhibits in vitro replication of a wide range of influenza viruses, including those resistant to currently available drugs.

scholarly journals Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium

2006 ◽  
Vol 80 (19) ◽  
pp. 9896-9898 ◽  
Author(s):  
Eva Böttcher ◽  
Tatyana Matrosovich ◽  
Michaela Beyerle ◽  
Hans-Dieter Klenk ◽  
Wolfgang Garten ◽  
...  
Keyword(s):  
Serine Proteases ◽  
Mammalian Cells ◽  
Mdck Cells ◽  
Influenza Viruses ◽  
Human Respiratory Tract ◽  
Proteolytic Activation ◽  
Mammalian Expression ◽  
Human Airway Epithelium

ABSTRACT Host cell proteases that cleave the hemagglutinin (HA) of influenza viruses in the human respiratory tract are still not identified. Here we cloned two human type II transmembrane serine proteases with known airway localization, TMPRSS2 and HAT, into mammalian expression vector. Cotransfection of mammalian cells with plasmids encoding HA and either protease resulted in HA cleavage in situ. Transient expression of either protease in MDCK cells enabled multicycle replication of influenza viruses in these cells in the absence of exogenous trypsin. These data suggest that TMPRSS2 and HAT are candidates for proteolytic activation of influenza viruses in vivo.

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