Inversed air-liquid interface models for co-culture of airway epithelium with immune cells

Background

Respiratory tract infections are common in livestock and humans and can cause severe disease, however, tools to study respiratory pathogens in vitro are limited. Primary or organoid-based airway epithelial cells (AECs) cultured at air-liquid interface are increasingly used for studying the pathogenesis of respiratory virus infections. These models consist of a well-differentiated stratified epithelium containing basal, mucus-forming and ciliated cells, with a top layer sealed by tight junctions. At Wageningen Bioveterinary Research (WBVR) such AEC models for cattle and chicken were recently developed and used to study relevant respiratory pathogens such as avian influenza virus (AIV) and bovine respiratory syncytial virus (BRSV). However, a major shortcoming of such in vitro models is the absence of immune cells, which are an important component of the submucosa of the respiratory epithelium in vivo. In recent years “upside-down” air-liquid interface models have been described, in which an air-liquid interface is created with the epithelium oriented downward. In this model, the immune cells can be added to the top compartment, where gravity will bring them in close contact with the basolateral side of the epithelium.

The project

Our objective is to develop upside-down models for bovine and chicken AECs. In the chicken model we will study the impact of different types of (innate) immune cells, including natural killer cells (NKs), dendritic cells (DCs) and monocytes on infection with avian influenza virus (AIV). In the bovine model we will study the impact of immune cells such as primary bovine neutrophils on infection with bovine respiratory syncytial virus (BRSV).