Ex vivo adenovirus-mediated gene transfer and immunomodulatory protein production in human cornea

Function of indoleamine 2,3‐dioxygenase in corneal allograft rejection and prolongation of allograft survival by over‐expression

“…The endothelial cells, which replicate at low rates in humans [33], are the critical target for intraocular allogeneic effector cells in rejection [34]. For these reasons, but also on account of the accessibility of the endothelium and feasibility of storage of excised donor corneas ex vivo for extended periods, corneal endothelium is an appealing target for gene-based approaches to immunomodulation and protection [35][36][37]. In these studies we used the lentivirus EIAV as the vector for gene transfer, on account of longer term transgene expression and minimal immunogenicity seen with this vector [38,39].…”

“…As an additional negative control, we used virus-free medium. Medium was exchanged with full medium after 1 h. Gene transfer to full-thickness corneas was carried out as previously described [35,54], corneal specimens were incubated in 250 lL of the virus preparation at an MOI of 100 for 60 min prior to transplantation. MOI was estimated based on the known density of corneal endothelial cells in untreated specimens [55].…”

Adenoviral reporter gene transfer to the human trabecular meshwork does not alter aqueous humor outflow. Relevance for potential gene therapy of glaucoma

“…of Av1LacZ4, 14 but not when rabbits were injected at lower concentrations (unpublished). In ex vivo cornea culture studies from humans, rabbits or rats, 18,19,35 where lower concentrations were used, the authors did not observe swelling or detachment. Although their cultures were not infected under perfusion pressure, treated corneas were able to keep corneal clarity after transplantation in rabbits.…”

Co-injection of adenovirus expressing CTLA4-Ig prolongs adenovirally mediated lacZ reporter gene expression in the mouse retina

“…19 This protein has previously been shown to be functionally active, as determined by binding to B7.1 expressing transfectants. 19 A series of BALB/c mice was subretinally injected with a mixture of AdCTLA and AdRL, an E1-deleted adenoviral vector containing a lacZ reporter gene also driven by an RSV promoter. The surgical procedure was carried out as previously described.…”

“…A number of vectors, including those based on herpes simplex virus (HSV), 2 lentivirus 3 and adeno-associated virus (AAV) [4][5][6][7] have been used for ocular gene transfer but, to date, the most widely reported system is that based on adenoviral (AV) vectors. [8][9][10][11][12][13][14][15][16][17][18][19] Studies in either the rabbit or more often the mouse, using these AV vectors carrying a lacZ reporter gene, have demonstrated that cells in the anterior segment, including the corneal endothelium, iris pigment epithelium, ciliary body, trabecular meshwork and Schlemms canal, may be efficiently transduced by intra-cameral injection. Subretinal injection is required for efficient targeting of the RPE and also results in the transduction of a small proportion of photoreceptors.…”