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Brief description/objective

The human airway epithelium represents the entry port of many human respiratory viruses, such as respiratory syncytial virus (RSV), influenza A viruses (IAV), and human coronaviruses (HCoV). It represents the interface between the external world and mucosal tissues and serves as a first biological and physical barrier to pathogen infection. We have recently established the cultivation of primary human airway epithelia (HAE) cells that enable studies of human respiratory virus infections in a culture system that morphologically and functionally resembles human airways in vivo. The pseudo-stratified HAE cell layer contains basal, secretory, columnar and ciliated cell populations, and generates mucus. Therefore, this in vitro system recapitulates many aspects of the human airway epithelium, namely (i) the presence of well defined cell types of the human airway epithelium, and (ii) physical/biochemical barriers, such as the mucus layer.
In the current project we will analyze the secretome of HAE cell layers following human respiratory virus infection. We will use the newest generation of mass spectrometry-based discovery tools to determine the secretome of infected HAE cultures following infection with RSV, IAV and HCoV. HAE cultures represent truly polarized airway cell layers, and we hypothesize (i) that host proteins secreted apically (i.e. towards the mucus) will be very different compared to host proteins secreted basolaterally (towards the medium; i.e. towards the mucosal tissue in vivo). We further hypothesize (ii) that respiratory virus infection will reveal a common pattern within the secretome that is generally induced by viral infections, as well as a distinct pattern that is characteristic for each specific respiratory virus.

We expect that our studies will provide a first glance concerning the magnitude of changes within the secretome of HAE cells following respiratory virus infection. Furthermore, our studies will provide a starting point towards the identification of candidate molecules that may have antiviral function within the mucus layer, and they will provide important insights into the pattern of basolaterally secreted chemokines and cytokines that are decisive to shape inflammatory and adaptive immune responses.