Applying Human and Porcine Primary Cell Air-Liquid Interface Cultures to Study Viral Coinfections

Across the globe, coronaviruses and influenza viruses co-circulate within human and pig populations. Respiratory viral coinfections within both populations present a global health concern, contributing to morbidity, mortality, and imposing economic burdens [1]. However, coinfections remain poorly understood and vastly understudied. Respiratory viruses are obligate intracellular pathogens which have evolved to exploit a myriad of host factors to support all stages of their replication cycle and most commonly infect airway epithelial cells as their primary targets [2,3]. To enable the study of coinfections, we require a suitable in vitro model of the respiratory epithelium [4]. To mimic the in vivo physiology of the respiratory epithelium we propose culturing human and porcine primary airway epithelial cells (PAEC) at the air-liquid interface (ALI). Furthermore, we aim to develop macrophage co-cultures or sub-cultures within the PAEC-ALI model, enabling us to model the cross-talk between epithelial and immune cells. Initial experiments will characterise mono- and co-infections, target cells, and cell responses. We will use RNA- and single-cell RNA sequencing to explore respiratory viral coinfections to identify candidate genes involved in exacerbation or reduction of viral infection. Validation of select candidate genes aims to identify genetic susceptibility or resilience markers or targets for antiviral intervention.