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NA Loewen, MP Fautsch, CK Bahler, DH Johnson, EM Poeschla; Effects of FIV-Based Lentiviral Vector Transduction of Trabecular Meshwork on Aqueous Outflow Facility and In Vivo Monitoring of Expression in Cats . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3960.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine the effects of feline immunodeficiency virus-based lentiviral vectors (FIV vectors) on outflow facility of cultured human anterior segments and to evaluate these vectors in the anterior chambers of living cats. Methods: FIV.lacZ vectors, which we have previously shown effectively transduce a beta-galactosidase marker gene into the TM of cultured human anterior segments, and new FIV vectors encoding eGFP (FIV.egfp), were used. 3 pairs of human donor eyes were injected with 1x10^8 TU (transducing units) of FIV.lacZ (right eyes) and with a control vector (FIV.lacZ.mock, left eyes). 3 additional pairs were injected with 1x10^8 TU FIV.egfp (right eyes only). Intraocular pressure was continuously recorded for 5 days and transduction efficiency was determined as the percentage of transduced cells in all quadrants. The right eyes of 2 cats were injected with 1x10^8 TU of FIV.egfp and expression levels in the living animals were rated using gonioscopy and biomicroscopy on a scale from 0 - 4. Results: Both FIV.egfp and FIV.lacZ resulted in high-level transduction of TM (79 ± 15% and 82.4 ± 3.6% of cells respectively) without cell loss compared to control eyes. Transduction caused only a transient 30% ± 22% (p = 0.02) decrease of outflow facility in cultured human anterior segments, lasting about 2 days. Facility in control eyes remained unchanged. 7 and 25 days after injection of cats 1 and 2 respectively, gonioscopically visualized eGFP expression was level 3-4 and localized to the TM; longer follow-up and correlative histological examination will be performed. Conclusion: High-level eGFP and beta-galactosidase expression can be achieved in TMs of cultured human eyes, with only small and transient changes in outflow facility. eGFP can be transduced into feline TMs with FIV vectors and can be visualized non-invasively in vivo.
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