Empirical Study Of Genetic Transmission To The Eye With Lentiviral Vector

ObjectiveTo survey the infection efficiency of exogenous gene transferring to RPE cells by Lentiviral vector, and an influence of virus titer, period of infection and toxic action on the cells as well. To estimate the effects and feasibility of GFP and Endostatin gene transferring to eyes of mice. To approach the potentiality of inhibiting the growth of retina neovascular and to provide prophase foundation of the treatments of ocular neovascular diseases by genetic transmission.MethodsThe Lentiviral vector with exogenous gene(GFP and ES) and two different promoters(CMV and mRhodopsin) infected RPE cells in vitro and retina via intravitreous injection in vivo. The infection efficiency was evaluated by the ratio and fluorescence intensity of RPE cells expressing GFP. Toxic action of Lentiviral vector to RPE cells was known through transmission electron microscope(TEM) and MTT tests. The transmission effects in eyes were estimated by the expression of fluorescence in mice retina.To observe the appearance and quantity of retina neovasculars by constructing the mice ROP models.ResultsThe quantity of RPE cells increased and fluorescence intensity enhanced with the increasing of viral titer and period time of infection. Also RPE cells expressed Endostatin increasingly with viral titer raising. The viral vectors did not have obvious impact on proliferation and ultramicrostructure of RPE cells. Intravitreal injection of Lentival vector with GFP gene made it possible to express the gene in mice retina. In ROP models nonperfusion areas and more retinal neovasculars than that of control groups could be seen in mice retina.ConclusionsExogenous gene could be efficiently and specially transferred into RPE cells by Lentivirual vectors, which depended on dose of virus and the period of time of infection. The vectors within some viral titer did not have obvious impact on proliferation and ultramicrostructure of RPE cells. It was efficient to transfer exogenous gene to retina by intravitreal injection of Lentival vectors. The success of formation of mice ROP models showed potentiality for vascular growth inhibiting factor transferring to the eye.