FIG 5.

Immunohistochemistry using rabbit TGs following corneal delivery of ssAAV8-GFP-Y733 and a KOS/62 demonstrates colocalization of both the vector and HSV-1 in sensory neurons. Serial sections of TG from rabbits inoculated with ssAAV8-GFP-Y733 and KOS/62 were used to demonstrate that the vector and HSV-1 colocalize in neurons. Serial sections of <5 μm are presented at magnifications of ×10. IHC to detect the presence of KOS/62 was done using a mouse anti-β-galactosidase primary antibody (Abcam), followed by a biotinylated HRP secondary antibody (Vector Laboratories). To detect AAV8-GFP in neurons, sections were incubated with mouse anti-GFP antibody (Abcam), followed by incubation with a biotinylated HRP secondary antibody (Vector Laboratories). IHC was done to detect either HSV-1 or AAV8 using serial sections to demonstrate that neurons harbor both the vector and the virus. Control slides were prepared from coinfected rabbit TG, subjected to IHC with no primary antibody.