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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-CellTM 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.

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