Neuroinflammatory consequences of rhinovirus infection in the human airway

Abstract

Rhinovirus (RV) is the leading cause of exacerbations of airway disease, during which the airways become hypersensitive and manifests clinically with troublesome bouts of cough. The mechanism responsible for respiratory virus induced cough in humans is unknown. This research aimed to determine whether RV directly infects and alters neural function or exerts its effect indirectly via the release of inflammatory mediators from infected airway epithelium.

A human sensory neuronal model was derived from differentiation of human dental pulp stem cells towards a neuronal phenotype, termed peripheral neuronal equivalents (PNEs). PNEs were infected for 2, 6, 24, 48 and 72 hrs with major group RV-A16, or minor group RV-A1b, at a range of multiplicities of infection (MOI) (0.01 – 100). Primary bronchial epithelial cells (PBEC) from chronic cough, COPD and healthy patients, were infected for 24 hrs with RV-A16 and at MOI 1.

PNEs expressed intracellular adhesion molecule 1 (ICAM-1), the entry receptor of major group RV. Viral titrations, Western Blotting (WB), immunocytochemistry (ICC) and flow cytometry confirm RV-A16 entry and replication in PNE when infected at MOI 1 for 24, 48 and 72 hours. RV-A1b infection at the same MOI and infection times, failed to infect PNE cells to detectable levels. A human antibody array indicated upregulation of several cytokines following RV-A16 infection, most notably IL-1β at 24 hours post infection (h.p.i) and IL-4 at 48 h.p.i. Viral titrations, WB and ICC confirmed RV-A16 replication in PBEC at MOI 1 at 24 h.p.i. Significant increases IL-1β were observed in chronic cough and COPD PBEC infected at MOI 1. Treatment of PNE with IL-1β for 24 hrs resulted in TRPA1 channel sensitivity in response to TRPA1 agonist, cinnamaldehyde.

We report for the first time the susceptibility of an airway neuronal model to RV infection. Findings confirm RV-A16 entry and replication in PNE, as well as a pro-inflammatory cytokine release.

Thesis embargoed until 31 July 2027.

Date of Award	Jul 2022
Original language	English
Awarding Institution	Queen's University Belfast
Sponsors	BREATH project, supported by the European Union’s INTERREG VA Programme
Supervisor	Lorcan McGarvey (Supervisor), Fionnuala Lundy (Supervisor) & Lorraine Martin (Supervisor)