Neuroinflammatory Consequences of Rhinovirus Infection in Human Epithelial and Neuronal Models

Rhinovirus (RV) is the leading cause of exacerbations of lung disease. A sensory neuronal model, derived from human dental pulp stem cells and differentiated into peripheral neuronal equivalents (PNEs), was used to examine RV's effects on airway sensory nerves. We investigated whether RV can directly infect and alter PNEs or whether it exerts effects indirectly via the release of mediators from infected epithelial cells.

PNEs or primary bronchial epithelial cells (PBECs) were infected with the RV-A16 strain. Viral replication was confirmed by viral titration assays, immunofluorescence (IF) for the double-stranded RNA (dsRNA) replication intermediate and western blotting (WB). RNA sequencing was used to determine transcriptomic changes in PNEs, and inflammatory responses were assessed by inflammatory microarray. Calcium mobilisation assays were used to investigate the effect of interleukin-1β (IL-1β) on PNE transient receptor potential (TRP) A1 channel responses.

Viral titrations, WB and IF confirm RV-A16 entry and replication in PNEs and PBECs. Gene signatures associated with antiviral immune responses, sensory neuropathies and N-Methyl-D-aspartic acid (NMDA) receptor activity were upregulated in RV infected PNEs. Several cytokines were increased from PNEs and PBECs following RV infection, most notably IL-1β. Treatment of PNEs with IL-1β resulted in heightened TRPA1 channel sensitivity.

We report the suitability of an airway neuronal model for the study of the direct effects of RV infection on nerves. RV-induced release of IL-1β from airway epithelium heightens neuronal TRPA1 responses suggesting a mechanism for virus-induced cough hypersensitivity.