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Q. Li, J. Pang, R. Miller, A. Dinculescu, V. Chiodo, W. W. Hauswirth; Intraocular Route of AAV Vector Administration Defines Immune Response and Therapeutic Potential. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4759.
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Safety and efficiency are critical for successful gene therapy. Adeno-associated viral (AAV) vectors are commonly used for gene transfer in both human and animal studies. However, administration of AAV-vectors can lead to development of neutralizing antibodies against the vector capsid, thus decreasing the efficiency of therapeutic gene transfer and preventing effective vector re-administration. We investigated immune responses to different routes of ocular administration and re-administration of AAV vectors, and the effect of previous exposure of AAV vector in one eye on the transduction efficacy of subsequent intra-ocular AAV-mediated gene delivery to the partner eye.
We tested two vector systems. One contains a cDNA encoding a secreted Pigment Epithelial Derived Factor (PEDF) cDNA under the control of a CMV enhancer/chicken β-actin promoter (CBA) (AAV2-CBA-PEDF) and was tested in a murine model of laser induced choroidal neovascularization (CNV). The other vector contains a cDNA encoding the intracellular reporter protein GFP under the control of the same promoter (AAV2-CBA-GFP). Animals were divided into groups, and received sequential injections at different combinations of either intravitreal or subretinal routes. CNV was evaluated by fluorescein angiographic choroidal flat-mount image analysis. The expression of GFP was analyzed in retinal sections by direct fluorescence imaging. Antibodies against AAV2 capsid were analyzed by ELISA using serum samples collected prior to injection and different time points after the injection.
Different ocular compartments respond to AAV administration differently. Intravitreal administration of AAV vectors, which results in transduction of inner retina (primarily retinal ganglion cells), generates humoral immune response against AAV capsid that blocks vector expression upon re-administration via the same route into the partner eye. In contrast, it has no effect on vector re-administered into the sub-retinal space of the partner eye. Additionally, subretinal administration of vector does not trigger any humoral immune response against AAV capsid, and has no effect on subsequent administration of vector either intravitreally or subretinally into the partner eye.
These findings have important clinical implications for the design of AAV-mediated ocular gene transfer for retinal diseases, particularly if both eyes require sequential treatment.
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