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TM Issa el Samman, DR Saban, IA Elder, TB Nguyen, MB Grant, AM Timmers; Prolongation of Recombinant Adenoviral Expression Through Modulation of the Innate Ocular Immunity . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2733.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: The innate immune inflammatory response is a major pathway in the activation of cellular-mediated clearance of ocular infection. In this study we tested indomethacin, a powerful non-steroidal anti-inflammatory drug, to inhibit this inflammatory response for the prolongation of subretinally delivered recombinant adenoviral passenger gene expression. Methods: Sprague-Dawley rats (males of 200g) received indomethacin treatment in their drinking water (1.2 mg/day/kg in 0.01M phosphate buffer, pH 7.2). First, we tested the effect of indomethacin treatment on water intake, weight, and the histology of vital organs. Next, another set of rats received 4*107 Ad-CMV-GFP infectious particles by injection into the subretinal space of the right eyes, leaving the contralateral eyes uninjected. Rats were divided in 3 groups - a control group which received no treatment, a short-term group which received treatment for 2 weeks post-injection, and a long-term group which received treatment from injection until sacrifice. Full-field scotopic ERG at 3-week intervals assessed electrophysiological changes in the retinas of all groups. Overnight dark-adapted rats were exposed to white-light flashes with an increasing intensity of -4.4 to 0.4 log cd.s.m-2.After sacrifice, eyecups were cryosectioned to analyze GFP expression. Cresol violet staining was done to assess inflammatory responses in the eyecups. Results: Maintenance of rats on indomethacin did not affect weight increase, water intake, nor did it induce pathology of vital organs including heart, kidney, spleen, lung and liver. Furthermore, A-wave and B-wave amplitudes of indomethacin treated animals were identical to non-treated control animals. In the rats injected with Ad-CMV-GFP, the injected eyes of all three groups showed an immediate 30% decrease in ERG amplitudes without substantial recovery. Cresol violet staining confirmed the suppression of inflammation in short and long-term groups. GFP expression in our control group lasted approximately 6 weeks, while both short and long-term groups showed expression up to 21 weeks. Conclusion: The loss of ERG signal as a result of adenoviral injection remained suppressed even though inflammation was inhibited. Our results also illustrate that the inhibition of the innate inflammatory response allows for the prolongation of subretinally delivered recombinant adenoviral passenger gene expression.
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