Abstract
Purpose :
To investigate the response to nucleic acid stimulation in the membrane-associated mucin expression in immortalized corneal and conjunctival epithelium.
Methods :
Immortalized human corneal epithelial cells (HCLE) and human conjunctival epithelial cells (HCjE) were cultured on 12-mm Transwell filters (n=12) at a density of 4x105 cells/cm2. The cultured cells were then stimulated with 25µg/ml of polyinosinic:polycytidylic acid [Poly(I:C)], an analog of viral double-stranded RNA produced during viral replication. Transepithelial electrical resistance (TER) was then measured using EndOhm electrodes (World Precision Instruments). After 6, 12, and 24 hours of exposure to Poly(I:C), the expressions of membrane-associated mucins (MUC1, MUC4, and MUC16) were analyzed by Western blotting. Immunoreactive bands were visualized by chemiluminescence, and densitometry analysis was then performed.
Results :
Poly(I:C) challenge increased the TER in a time-dependent manner. After 6 hours and 12 hours of Poly(I:C) exposure, no change was observed in all 3 membrane-associated mucins. After 24 hours, membrane-associated mucins were increased.
Conclusions :
The findings of this study show that Poly(I:C) challenge, which mimics a viral infection, increased the barrier function of ocular surface epithelia by increasing the membrane-associated mucin expression. Thus, we theorize that the increased barrier function must be a kind of host defense reaction to viral infection.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.