July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Nucleic acid stimulation increases galectin-3 expression in immortalized corneal and conjunctival epithelium
Author Affiliations & Notes
  • Yuriko Ban
    Ophthalmlogy, Kyoto Chubu Medical Center, Nantan, Kyoto, Japan
    Ophthalmlogy, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Naoko Miyagi
    Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan
  • Chie Sotozono
    Ophthalmlogy, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Shigeru Kinoshita
    Frontier Medical Science and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Footnotes
    Commercial Relationships   Yuriko Ban, None; Naoko Miyagi, None; Chie Sotozono, None; Shigeru Kinoshita, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 3842. doi:
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      Yuriko Ban, Naoko Miyagi, Chie Sotozono, Shigeru Kinoshita; Nucleic acid stimulation increases galectin-3 expression in immortalized corneal and conjunctival epithelium. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3842.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : On the ocular surface, galectin-3, an endogenous carbohydrate-binding protein, colocalizes with membrane-associated mucins and contributes to the stabilization of the glycocalyx barrier. In this study, we investigated the response to nucleic acid stimulation in galectin-3 expression in immortalized corneal and conjunctival epithelium.

Methods : Immortalized human corneal epithelial (HCLE) cells and human conjunctival epithelial (HCjE) cells 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 exposure to Poly(I:C), the expressions of galectin-3 mRNA were analyzed by real-time polymerase chain reaction. The expressions of galectin-3 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-, 12-, and 24-hours exposure to Poly(I:C), galectin-3 mRNA increased (p<0.01). After 6 hours and 12 hours, no change was observed in galectin-3 expression, yet after 24 hours, galectin-3 expression increased (p<0.01).

Conclusions : The findings of this study show that Poly(I:C) challenge increases galectin-3 expression. We previously reported that Poly(I:C) challenge also increases the expression of membrane-associated mucins (ARVO 2016 & 2017). Our findings revealed that Poly(I:C) challenge, which mimics a viral infection, increases the barrier function of ocular-surface epithelia. 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 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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