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M. Di Nolfo, A. Lombet, C. Blondin, W. Rostène, C. Baudouin, F. Brignole; Screening Chemokine Receptors on a Conjunctival Cell Line by Intracellular Calcium Measurement . Invest. Ophthalmol. Vis. Sci. 2003;44(13):704.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose : Chemokines are peptides with chemotactic properties which confer them an important role in the inflammatory reaction. We have sought for a wide variety of chemokine receptors on a human conjunctival cell line (Chang cells). Methods: Conjunctival cells from Chang cell line were investigated at baseline and after incubation with IFN-g at different concentrations (20 and 200 IU/ml) for 72 hours. These cells with or without IFN-g stimulation were stimulated by various human recombinant chemokines (six CC chemokines, SDF-1, RANTES, MIP-1a, Eotaxin, TARC, and MCP-1 ; two CXC chemokines, IP-10, IL-8 and the only CX3C chemokine, Fraktalkine). Cells were observed with a station Quanticell 700 which permits the monitoring of the cytosolic calcium during stimulation by chemokines. If the chemokine receptor is present and functional, then is observed a variation in cytosolic calcium. Three experiments were performed for each chemokine at least. Results: The receptor for SDF-1 was present and functional on our cellular model with 100 % of cells presenting calcium variation. Receptors for RANTES and Fraktalkine were present at a lower level with respectively 75 and 50% of the cells showing variation of cytosolic calcium after stimulation. There was no difference between cells at baseline and after treatment with IFN-g for these chemokines. We observed a mild effect of IP-10 on our cells when treated with 200 IU/ml of IFN-g. Conclusions: Our results suggest that CXCR-4 is present on Chang at baseline. These preliminary findings need more investigation such as in vivo expression of CXCR-4 and SDF-1 in the ocular surface. This chemokine is implicated in angiogenesis. The description of this system could be interesting in the understanding and treatment of corneal neovascularization.
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