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RL Hurwitz, T Mahoney, J-R Lin, C Hurwitz, MY Hurwitz; Adenoviral Serotype Influences the Efficiency of Adenoviral-Mediated Transgene Delivery for Gene Therapy . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4621.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To compare the efficiency of adenoviral serotype 5 vectors to chimeric adenoviral serotype 5 vectors expressing adenoviral serotype 35 fibers in transgene expression for ocular gene therapy Methods: Adenoviral 5 constructs containing a green fluorescent reporter gene (AdV5-GFP) were compared to similar constructs using a chimeric adenovirus expressing fiber proteins derived from adenovirus serotype 35 (AdV5/F35-GFP). Y79 retinoblastoma cells were incubated with either adenoviral construct and the expression of the reporter protein was compared using quantitative fluorescence and FACS analysis. The adenoviral receptor CAR was measured by FACS using a monoclonal antibody to human CAR. Results: CAR was confirmed to be present on Y79 cells in amounts comparable to those in other cell lines known to be transduced by AdV5. The AdV5/F35 vector was more effective by one order of magnitude viral concentration than the AdV5 vector in transducing the retinoblastoma cells. The maximum amount of GFP as measured by fluorescence expressed after transduction by the AdV5/F35 vector was more than 2-fold greater than the maximum amount of GFP expressed after transduction by the AdV5 vector. Conclusion: An order of magnitude less AdV5/F35 vector could induce the expression of twice the amount of GFP protein than that mediated by the AdV5 vector. AdV5/F35 may be superior to AdV5 as vector for some ocular gene therapy applications.
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