June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Galectin-1 and -3 as prosprective biomarkers in keratoconus patients
Author Affiliations & Notes
  • Rossen M Hazarbassanov
    Ophthalmology and Visual Sciences, Paulista School of Medicine, Federal University of Sao Paulo, Sâo Paulo, SP, Brazil
  • Frans Eberth Costa Andrade
    Morphology and Genetics, Paulista School of Medicine, Federal University of Sao Paulo, Sâo Paulo, SP, Brazil
  • Joyce Covre
    Ophthalmology and Visual Sciences, Paulista School of Medicine, Federal University of Sao Paulo, Sâo Paulo, SP, Brazil
  • Myrna Serapião dos Santos
    Ophthalmology and Visual Sciences, Paulista School of Medicine, Federal University of Sao Paulo, Sâo Paulo, SP, Brazil
  • Mauro Silveira de Queiroz Campos
    Ophthalmology and Visual Sciences, Paulista School of Medicine, Federal University of Sao Paulo, Sâo Paulo, SP, Brazil
  • Jose Alvaro Pereira Gomes
    Ophthalmology and Visual Sciences, Paulista School of Medicine, Federal University of Sao Paulo, Sâo Paulo, SP, Brazil
  • Cristiane Damas Gil
    Morphology and Genetics, Paulista School of Medicine, Federal University of Sao Paulo, Sâo Paulo, SP, Brazil
  • Footnotes
    Commercial Relationships   Rossen Hazarbassanov, None; Frans Eberth Costa Andrade, None; Joyce Covre, None; Myrna Serapião dos Santos, None; Mauro Campos, None; Jose Gomes, None; Cristiane Damas Gil, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 3909. doi:
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      Rossen M Hazarbassanov, Frans Eberth Costa Andrade, Joyce Covre, Myrna Serapião dos Santos, Mauro Silveira de Queiroz Campos, Jose Alvaro Pereira Gomes, Cristiane Damas Gil; Galectin-1 and -3 as prosprective biomarkers in keratoconus patients. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3909.

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

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Abstract

Purpose : To evaluate the expression of galectin-1 (Gal-1) and -3 (Gal-3) in the keratoconus (KC) diagnosed patients and riboflavin and ultraviolet light effect on human keratocytes cultivated in vitro.

Methods : Tear fluid and conjunctival impression cytology specimens from control and KC patients with and without atopy (n = 10/group) were used to evaluate Gal-1 and Gal-3 expression by immunofluorescence and ELISA. Primary human keratocytes were isolated by digestion in collagenase from surgically removed corneas of five normal human corneal buttons and cultured in DMEM/Ham's F12 medium supplemented with 2 % fetal bovine serum. These cells were evaluated under two experimental conditions: control and submitted to the application of UVA light and riboflavin 0.1% for 30 minutes called cross-linking (CXL). After 24 hours of CXL, expression of Gal-1 and Gal-3 in the and their supernatants was performed using immunofluorescence and ELISA; multiplex assay was also done to detect inflammatory biomarkers in supernatants. Student’s t-test was used for statistical analysis.

Results : Atopic KC patients exhibited increased levels of Gal-1 in the tear fluids (14.5±3 ng/mL, p>0.05) and epithelial cells of bulbar conjunctiva (130±3 arbitrary units, p<0.001) compared to control (9.8±0.5 and 87±1.6, respectively). On the other hand, non-atopic KC patients exhibited increased levels of Gal-3 in the tear fluid (70±16 ng/mL, p<0.05), but decreased endogenous expression in conjunctival epithelial cells. In in vitro experiments, keratocytes exhibited intense nuclear Gal-1 expression while Gal-3 was localized especially in the cytoplasm and both were strongly diminished after 24 hours of CXL. Further, CXL induced significant release of Gal-1 in the cell supernatants (116±18 ng/mL, p<0.05) and decreased inflammatory biomarkers as IL-6, IL-8, MMP-2 and MMP-9. Gal-3 levels was not detected in the keratocyte supernatants

Conclusions : Gal-1 and Gal-3 represent new interesting biomarkers as revealed by their different expression patterns in atopic and non-atopic KC patients. CXL has immunosuppressive effect on keratocytes by reducing the release of cytokines and MMPs and increased anti-inflammatory protein Gal-1.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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