June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Role of galectin-3 in dry eye disease
Author Affiliations & Notes
  • Yuichi Uchino
    Schepens Eye Research Institute and Massachusetts Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA
  • Jerome Mauris
    Schepens Eye Research Institute and Massachusetts Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA
  • Julia Dieckow
    Schepens Eye Research Institute and Massachusetts Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA
  • Ashley Woodward
    Schepens Eye Research Institute and Massachusetts Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA
  • Francisco Amparo
    Schepens Eye Research Institute and Massachusetts Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA
  • Reza Dana
    Schepens Eye Research Institute and Massachusetts Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA
  • Flavio Mantelli
    Ophthalmology, Campus Bio-Medico, University of Rome, Rome, Italy
  • Pablo Argueso
    Schepens Eye Research Institute and Massachusetts Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA
  • Footnotes
    Commercial Relationships Yuichi Uchino, Santen Pharmaceutical (F); Jerome Mauris, None; Julia Dieckow, None; Ashley Woodward, None; Francisco Amparo, None; Reza Dana, Allergan (C), Alcon (C), Bausch & Lomb (C), Eleven Bio (I), GSK (F), Novabay (C), Revision Optics (C), Novaliq (C), RIgel (F); Flavio Mantelli, None; Pablo Argueso, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2666. doi:https://doi.org/
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      Yuichi Uchino, Jerome Mauris, Julia Dieckow, Ashley Woodward, Francisco Amparo, Reza Dana, Flavio Mantelli, Pablo Argueso; Role of galectin-3 in dry eye disease. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2666. doi: https://doi.org/.

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

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Abstract

Purpose: Galectin-3, a multimeric carbohydrate-binding protein expressed by the ocular surface epithelia, provides barrier function under physiological conditions through interaction with transmembrane mucins on the apical glycocalyx. Here, we investigated whether galectin-3 is altered on the ocular surface of dry eye patients and/or modulates cytokine receptor activation in corneal epithelial cells.

Methods: Galectin-3 in tear fluid collected from the inferior fornix of normal subjects (n=11) and patients with dry eye (n=20) was analyzed by quantitative Western blot with recombinant protein as calibration standards. siRNA targeting galectin-3 and scramble control were introduced into stratified cultures of human corneal-limbal epithelial (HCLE) cells by lipofectamine-mediated transfection. Stratified HCLE cell cultures were treated for 2 h with serum-free growth-media alone or with recombinant human IL-1β (10 ng/ml). Secreted IL-8, IL-1 receptor 1 (IL1R1), and GAPDH were quantified by Western blot.

Results: The levels of galectin-3 detected in normal tear samples were 0.12±0.14 ng/µg total protein (range, 0.00-0.41). In dry eye, there was a significant increase in galectin-3 content in tears (0.38±0.37 ng/µg total protein; range, 0.04-1.36). In vitro, we found that galectin-3 mediates inflammatory responses in corneal epithelial cells. As expected, IL-8 protein levels were increased in the culture media of control HCLE cells treated with IL-1β. However, IL-8 secretion was impaired in HCLE cells treated with IL-1β after galectin-3 siRNA knockdown, indicating that galectin-3 mediates IL1R1 activation. The amount of IL1R1 in total cell protein extracts was not affected by either IL-1β stimulation or galectin-3 knockdown.

Conclusions: Increased levels of galectin-3 in the tear fluid may reflect alterations of the ocular surface glycocalyx barrier in dry eye patients. Galectin-3 regulates IL-1-mediated inflammatory responses on the ocular surface epithelia, but the molecular mechanisms of this regulation remain to be elucidated.

Keywords: 486 cornea: tears/tear film/dry eye • 482 cornea: epithelium • 485 cornea: surface mucins  
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