Effect of human rhinovirus infection on airway epithelium tight junction protein disassembly and transepithelial permeability.

Rationale: No studies have assessed the effects of human rhinovirus (HRV) infection on epithelial tight junctions (TJs) and resultant barrier function.

Aim of the Study: To correlate viral infection with TJ disassembly, epithelial barrier integrity, and function.

Materials and Methods: Human airway epithelial cells were infected with HRV minor serotype 1B (HRV-1B) at various 50% tissue culture infectivity doses (TCID50) over 72 hours. HRV replication was assessed by quantitative-polymerase chain reaction (qPCR)while cell viability and apoptosis were assessed by proliferation and apoptotic assays, respectively. Protein expression of claudin-1, occludin, and zonula occludens protein-1 (ZO-1) was assessed using In-Cell^TM Western assays. Transepithelial permeability assays were performed to assess effects on barrie rfunctionality. RT² Profiler focused qPCR arrays and pathway analysise valuating associations between human TJ and antiviral response were performed to identify potential interactions and pathways between genes of interests.

Results: HRV-1B infection affected viability that was both time and TCID50 dependent. Significant increases in apoptosis and viral replication post-infection correlated with viral titer.Viral infection significantly decreased claudin-1protein expression at the lower TCID₅₀, while a significant decrease in all three TJ protein expressions occurred at higher TCID₅₀. Decrease in protein expression was concomitant with significant increases in epithelial permeability of fluorescein isothiocynate labeled-dextran 4 and 20 kDa. Analysis of focused qPCR arrays demonstrated a significant decrease in ZO-1 gene expression. Furthermore, network analysis between human TJ and antiviral response genes revealed possible interactions and regulation of TJ genes via interleukin (IL)-15 in response to HRV-1B infection.

Conclusion: HRV-1B infection directly alters human airway epithelial TJ expression leading to increased epithelial permeability potentially via an antiviral response of IL-15.

Authors: Kevin Looi, Niamh M. Troy, Luke W.Garratt, Thomas Iosifidis, Anthony Bosco, Alysia G. Buckley, Kak-Ming Ling, Kelly M. Martinovich, Elizabeth Kicic-Starcevich, Nicole C.Shaw, Erika N.Sutanto, Graeme R. Zosky, Paul J. Rigby, Alexander N. Larcombe, Darryl A.Knight, Anthony Kicic and Stephen M.Stick.

Published in Experiemental Lung Research in August 2016.