Purchase this article with an account.
J Hargitai, K Doi, J Kong, L Ivert, P Gouras; Lentiviral Transduction of Vascular Endothelial Growth Factor (VEGF) in Retinal Pigment Epithelium Causes Vascular Changes in Rabbit . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4626.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To study the effects of lentiviral transduction of VEGF in the retina. Methods: A lentiviral vector expressing the murine VEGF164(lentiCMVVEGF164) was generated by cotransfection of 293T cells. Cultured human fetal retinal pigment epithelial cells (hfRPE) were infected with the virus. The transgene product was detected by Western blotting. 20 microliters of lentivirus at concentrations of 5x108-109 IU/ml were injected into the subretinal space of pigmented rabbits. Control rabbits received a subretinal injection of saline or mock virus. The development of vascular changes were followed by fluorescein (FAG) and indocyanine green (ICG) angiography using a scanning laser ophthalmoscope (SLO). Results: In vitro results showed high levels of VEGF164 expression. In vivo dilation and tortuosity of retinal vessels were seen in the early phase of FAG at 14 days postinjection. Early phase ICG angiography revealed irregularities of the choroidal vasculature. Dot and plaque-like choriocapillary leakage was observed throughout the late phase of FAG and ICG angiography in all eyes. Leakage severity correlated with the amount of injected virus. These changes were maintained for at least 2 months. Rabbits are still being followed. High viral titers produced some disruptive change in the RPE. Vascular changes were not seen in rabbits injected with saline or mock virus. Conclusion: LentiCMVVEGF164 efficiently transduces RPE in vitro. Our previous results showed that a lentivirus expressing enhanced green fluorescent protein(EGFP) predominantly transduces RPE in vivo after subretinal delivery, implying that the transgene protein (VEGF) was produced by the RPE. Retinal and choroidal vascular changes were observed in all virus injected eyes. These vascular changes are characteristics of choroidal neovascularisation (CNV). Histology will determine the extent of the vascular pathology. Whether CNV can be produced by this approach and without any significant RPE damage remains to be determined.
This PDF is available to Subscribers Only