Abstract
Purpose :
Interferon β (IFN-β), a subtype of type I IFNs, has an anti-viral effect as part of a response to viral double-stranded RNA (dsRNA) through the Toll-like receptor (TLR) 3 and retinoic acid-inducible gene I (RIG-I) pathways. We previously evaluated the response of synthetic dsRNA stimulation on immortalized human corneal limbal epithelial (HCLE) and conjunctival epithelial (HCjE) cells and found an increased barrier function in those cells. In this study, we investigated the effect of type I IFNs on ocular-surface barrier function in cultured HCLE and HCJE cells.
Methods :
HCLE and HCjE cells were cultured on 12-mm Transwell® inserts (Corning®) and stimulated with 0ng/mL (control) and 0.25, 2.5, and 25ng/mL IFN-β, and transepithelial electrical resistance (TER) was measured before stimulation and every 3 hours up until 24-hours post stimulation. Tight junction [claudin (CL)-1, CL4, and CL7] and glycocalyx [mucin (MUC)-1, MUC4, and MUC16] protein expressions were then evaluated via real-time polymerase chain reaction (PCR), western blotting, and immunohistochemistry examinations.
Results :
At 24-hours post stimulation with 2.5 and 25ng/ml IFN-β, a significant TER increase of 25.0% and 34.6%, respectively, in the HCLE cells and 10.9% and 15.0%, respectively, in the HCjE cells was observed compared with that in the control cells (P<0.05). The TER increase began at 3 hours post stimulation, and remained up until 24 hours. Post stimulation with 2.5ng/mL IFN-β, real-time PCR (P<0.05), western blotting (P<0.05), and immunohistochemistry findings revealed increased expression of CL1, CL4, MUC1, MUC4, and MUC16, yet no change of CL7 expression, in both the HCLE and HCjE cells compared with the control cells.
Conclusions :
In this study, the evaluation of tight junction and glycocalyx protein expression in HCLE and HCjE cells post stimulation with IFN-β revealed that IFN-β stimulation increased the barrier function of those cells, thus illustrating that it plays a significant beneficial role of promoting an anti-viral effect on the ocular surface.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.