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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)
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.
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.
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.
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.
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