This gene therapy protocol has, for the first time, permanently reversed T1D with normal glucose tolerance in NOD mice and, as such, represents a novel therapeutic strategy for the treatment of T1D.
1 1 MONDAY 28 days in vivo in recipient sheep after orthotopic corneal allotransplantation of a transduced donor cornea. However, the kinetics of LV-mediated gene expression were slower than those achieved by comparable adenoviral vectors. Further, 24 hour transduction of ovine corneal endothelium with LV-IL10 resulted in mRNA levels that were logarithmically lower than those achieved by an adenoviral vector. Shortening the transduction period to 2 hours led to a further reduction in gene expression. The addition of 40 ;g/ml polybrene to the transduction mixture restored expression to levels comparable to those attained after a 24 hour transduction period, but at the cost of associated toxicity for corneal epithelium. Allografts transduced for 2-3 hours with LV-SV40-IL10 in the absence of polybrene were rejected at a median of 25 days (n=7) compared with 18 days in the combined sham-treated and LV-SV40-eYFP control group (n=14) (p=0.026). Vectors containing the PGK or EF promoter were not more effective than LV-SV40-eYFP in driving transgene expression in ovine corneas, but LV-CMV-eYFP generated signifi cantly better, faster expression at low multiplicity of infection. Interestingly, LV-mediated eYFP and IL10 expression was signifi cantly higher and more rapid in organ-cultured human corneas (the eventual target species of interest) than in ovine corneas. Conclusions: Ovine and human corneal endothelial cells were effi ciently transduced by the lentiviral vector. LV-SV40-IL10 prolonged ovine corneal allograft survival to a modest, albeit signifi cant extent. Species differences were apparent in LV-mediated transduction effi ciency, but promoter exchange signifi cantly improved transgene expression levels. Aims: Our aims were to examine graft survival and visual outcome after corneal transplantation and to identify risk factors for graft failure within a large, prospective national register. Methods: Records of 17,090 full-thickness (penetrating) corneal grafts were examined within a prospectively-maintained, national database, with annual archival follow-up extending from one to 22 years. Information was available on the recipient, donor, eye bank practice, surgical procedure, and on postoperative outcomes. Kaplan-Meier survival analysis was used to indicate variables of interest for Cox proportional hazards regression analysis. A model clustered by individual patient to control for inter-eye or inter-graft dependence was constructed to identify variables best predicting penetrating corneal graft failure. Visual acuity in the grafted eye was measured by best-corrected Snellen acuity (without pinhole). Results: The major indications for corneal transplantation were keratoconus (32%). bullous keratopathy (26%). failed previous graft (19%). corneal dystrophy (7%). and herpetic eye disease (4%). Probability of corneal graft survival was 0.87 at one year, 0.73 at 5 years, 0.62 at 10 years and 0.51 at 15 years. Variables predicting graft failure in multivariate analysis included transplant centre, location and volume o...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.