June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Influence of lipopolysaccharide on proinflammatory gene expression in human corneal, conjunctival and meibomian gland epithelial cells.
Author Affiliations & Notes
  • Afsun Sahin
    Department of Ophthalmology, Eskisehir Osmangazi University Medical School, Eskisehir, Turkey
    Schepens Eye Research Institute, Boston, Massachusetts, United States
  • Wendy R Kam
    Schepens Eye Research Institute, Boston, Massachusetts, United States
  • Raheleh Rahimi Darabad
    Schepens Eye Research Institute, Boston, Massachusetts, United States
  • Yang Liu
    Schepens Eye Research Institute, Boston, Massachusetts, United States
  • David A Sullivan
    Schepens Eye Research Institute, Boston, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Afsun Sahin, None; Wendy Kam, None; Raheleh Darabad, None; Yang Liu, None; David Sullivan, None
  • Footnotes
    Support  NIH grant EY05612, the Margaret S. Sinon Scholar in Ocular Surface Research fund, TUBITAK, and the Guoxing Yao Research Fund
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 3946. doi:
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      Afsun Sahin, Wendy R Kam, Raheleh Rahimi Darabad, Yang Liu, David A Sullivan; Influence of lipopolysaccharide on proinflammatory gene expression in human corneal, conjunctival and meibomian gland epithelial cells.. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3946.

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

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Abstract

Purpose : Lipopolysaccharide (LPS), a bacterial toxin, is known to stimulate leuokotriene B4 (LTB4) secretion by human corneal, conjunctival and meibomian gland epithelial cells. We hypothesize that this LTB4 effect represents an overall induction of proinflammatory gene expression in these cells. Our objective was to test this hypothesis.

Methods : We cultured immortalized human corneal (gift from Dr. James Jester, Irvine, CA), conjunctival (gift from Dr. Ilene Gipson, Boston, MA) and meibomian gland epithelial cells in the presence or absence of LPS (15 μg/ml) and ligand binding protein (150 ng/ml). Cells were then processed for the isolation of RNA and the evaluation of gene transcripts by utilizing Illumina BeadChips, background subtraction, cubic spline normalization and GeneSifter software.

Results : Our findings show that LPS induces a striking increase in proinflammatory gene expression in human corneal and conjunctival epithelial cells. These cellular reactions are associated with a significant upregulation of genes associated with inflammatory and immune responses (e.g. IL-1 beta, IL-8, and tumor necrosis factor), including those related to cytokine-cytokine receptor interactions, chemokine and Toll-like receptor signaling pathways, and chemotaxis. In contrast, with the exception of Toll-like signaling pathways, almost no proinflammatory ontologies were upregulated by LPS in human meibomian gland epithelial cells.

Conclusions : Our results support our hypothesis that LPS stimulates proinflammatory gene expression in human corneal and conjunctival epithelial cells. However, our findings also show that LPS does not elicit such proinflammatory responses in human meibomian gland epithelial cells.

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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