June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
HLA-DR expression/mobilization on the conjunctival epithelial cells exposed to hyperosmolarity and desiccative stress
Author Affiliations & Notes
  • Christophe Roubeix
    Institut de la Vision, Paris, France
    INSERM/UPMC Univ Paris 06, PARIS, France
  • Luisa Riancho
    Institut de la Vision, Paris, France
    INSERM/UPMC Univ Paris 06, PARIS, France
  • Christophe Baudouin
    Institut de la Vision, Paris, France
    Quinze-Vingts National Hospital of Ophthalmology, PARIS, France
  • Francoise Brignole-Baudouin
    Institut de la Vision, Paris, France
    Paris Descartes University, Toxicology Department, PARIS, France
  • Footnotes
    Commercial Relationships Christophe Roubeix, laboratoires THEA (R); Luisa Riancho, None; Christophe Baudouin, None; Francoise Brignole-Baudouin, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2129. doi:
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      Christophe Roubeix, Luisa Riancho, Christophe Baudouin, Francoise Brignole-Baudouin; HLA-DR expression/mobilization on the conjunctival epithelial cells exposed to hyperosmolarity and desiccative stress. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2129.

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

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Abstract

Purpose: To characterize the effects of two in vitro models of dry eye on the expression/mobilization of HLA-DR known as a biomarker of dry eye. We investigated the effect of hyperosmolarity (HO) and desiccation stress (DS) on the membrane expression and the induction of HLA-DR mRNA as a first step to inflammation in dry eye disease in correlation with other inflammatory biomarkers such as IL-6, IL-8.

Methods: Hyperosmolar (HO) and desiccative stresses (DS) were induced on conjunctival cells (Wong-Kilbourne-derived of Chang Conjunctiva, WKD). WKD were exposed either to NaCl-induced HO conditions (500mOsm) for 24h or to desiccating conditions by seeding them on transwell membranes (d: 2.4cm, pore 0.4 μm) and incubated in a drying chamber (relative humidity: 17±3%) for 6h after cell culture medium removal from the upper side of the transwell. We evaluated the effect of HO or DS on IL-6, IL-8 and HLA-DR mRNA expression by qRT-PCR. IL-6 and IL-8 were measured for each stress condition using ELISA assay. HLA-DR cell was investigated using flow cytometry and immunofluorescence analyses.

Results: Both HO and DS induced an increase of IL-6 and IL-8 mRNA in WKD cells and a production of IL-6 and IL-8 in the supernatant. HO and DS induced a mild increase of the HLA-DR mRNA synthesis and a translocation of the cytoplasmic HLA-DR to the cell membrane confirmed by flow cytometry when compared to basal conditions.

Conclusions: By using these two in vitro dry eye models, we confirmed the proinflammatory profile induced by hyperosmolar or desiccating conditions on conjunctival cells. These conditions could trigger HLA-DR translocation at the cell membrane without the presence of IFNγ released by immune cell infiltration. These observations revealed the existence of a preliminary step in the chronic dry eye inflammation process.

Keywords: 474 conjunctiva • 557 inflammation • 726 stress response  
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