Developing an in vitro approach to study transmission of respiratory viruses

A key area of current influenza research attempts to understand the genetic basis of transmissibility. We do not know why the particular swine influenza virus that emerged in Mexico in 2008/9 caused a pandemic where as other swine influenza viruses, to which humans are exposed, did not. However the large number of experiments currently performed with ferrets use small animal numbers and crude readouts, and do not adequately measure difference in transmissibility between different viruses that might have large epidemiological impacts. For example, the length of contagiousness is not measured. Instead of using many more sentinel animals, we propose to develop methodology whereby sentinel animals are replaced by cultures of human airway epithelium that recapitulate the innate barriers that respiratory viruses must overcome before they initiate infection.

The project will use the H1N1 2009 pandemic strain of influenza A virus as a model because we and others already have a body of results generated in ferrets to compare and validate the in vitro alternative. The idea is to allow infected donor ferrets to breathe into a chamber as if in a small room or confined space. We will ask whether the air breathed out will and initiate infection on a cell culture placed downstream. The days on which infected animals are contagious will be compared for different viruses to measure increases or decreases in transmission.

Once methodology is established, we will move to replacing the infected donor animals with infected airway cultures to assess the environmental conditions that affect transmission of different respiratory viruses through the air. Experiments showing influenza virus transmission to be optimal at dry and cold conditions are often used to explain the winter seasonality of influenza. But parainfluenza virus outbreaks occur in summer, so we aim to assess the effect of humidity and temperature on transmission of PIV and other respiratory pathogens through the air.

Elderfield RA, Koutsakos M, Frise R, Bradley K, Ashcroft J, Miah S, Lackenby A, Barclay WS (2015). NB protein does not affect influenza B virus replication in vitro and is not required for replication in or transmission between ferrets. Journal of General Virology 97(3): 593-601 doi:10.1099/jgv.0.000386

Elderfield RA, Parker L, Stilwell P, Roberts KL, Schepelmann S, Barclay WS (2015). Ferret airway epithelial cell cultures support efficient replication of influenza B virus but not mumps virus. Journal of General Virology 96(8): 2092-8 doi:10.1099/vir.0.000176

 

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Project grant

Status:

Closed

Principal investigator

Professor Wendy Barclay

Institution

Imperial College London

Grant reference number

NC/K00042X/1

Award date:

Feb 2013 - Jan 2016

Grant amount

£399,896