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J.-T. Chen; DcR3 Rescues IFN--Induced Apoptosis in Conjunctival Cells. Invest. Ophthalmol. Vis. Sci. 2007;48(13):551.
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The purpose of this study was to investigate the effect of interferon (IFN)γ on apoptosis and on expression of apoptotic and inflammation related proteins in conjunctival cells. Soluble receptor (DcR3) was also introduced into conjunctival cells to test possible protection against IFN-γ-induced apoptosis.
A human conjunctival cell line was incubated with different concentrations of IFN-γ (50U/ml to 500U/ml) for 1day, 3 days, 5 days and 7 days. After treatment, conjunctival cells were subjected to flow ccytometry. Apoptosis was measured with Annexin V and PI staining. Inflammation related protein HLA-DR and ICAM-1 as well as apoptotic related protein, fas and fasL were evaluated by flow cytometry. DcR3 cDNA was introduced into conjunctival cells through a selflimiting lentiviral vector. Protection of DcR3 on IFN-γ-induced apoptosis was determined via Annexin V and PI staining with flow cytometry.
IFN-γ (50U/ml to 500U/ml) induced prominent expression of ICAM-1 and minor expression of HLA-DR in a time and dose dependent manner. Apoptosis of conjunctival cells was also induced by IFN-γ in a dose and time dependent manner. Expression of Fas and FasL in IFN-γ-treated conjunctival cells were increased. DcR3 was successfully expressed in conjunctivaql cells through lentiviral gene transfer and IFN-γ-induced apoptosis was decreased upto 40% by the DcR3 expression.
In our study, IFNγ induced expression of inflammatory proteins and apoptotic proteins, as well as apoptosis of human conjunctival cells. Lentiviral gene transfer of DcR3 into conjunctival cells partially protected conjunctival cells against IFNγinduced apoptosis.
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