AIM: In this paper we compare the transduction efficiency, toxicity, and safety of retroviral vectors [equine infectious anemia virus (EIAV), human immunodeficiency virus-1 (HIV-1), human foamy virus (PFV] and adenovirus (Ad) for potential use in gene therapy of corneal endothelial cells. METHOD: Murine corneal endothelial cells were transduced with EIAV, HIV-1, PFV, and Ad, resulting in the overexpression of a green fluorescent protein (eGFP) transgene marker. The transduction efficiency was assessed by flow cytometry, while cytotoxicity and apoptosis rate were detected by annexin V/propidium iodide (PI) stain. RESULTS: Ad had the highest transduction efficiency with 99% of the cells expressing the transgene, followed by EIAV (95%), HIV-1 (75%), and PFV (43%). However, the high transduction efficiency of Ad also resulted in the highest apoptosis rate (25%) in the corneal endothelial cells. There was no detectable difference in the toxicity between PFV and HIV-1 (10%). EIAV transduction had the lowest cytotoxicity, with only 3% of the cells being annexin V/PI positive. CONCLUSION: Compared to other vectors EIAV exhibited high transduction efficiency combined with low toxicity to corneal endothelial cells. Therefore, it is a powerful tool for gene therapy applications in selected corneal endothelial diseases.
AIM: In this paper we compare the transduction efficiency, toxicity, and safety of retroviral vectors [equine infectious anemia virus (EIAV), human immunodeficiency virus-1 (HIV-1), human foamy virus (PFV] and adenovirus (Ad) for potential use in gene therapy of corneal endothelial cells. METHOD:Murine corneal endothelial cells were transduced with EIAV, HIV-1, PFV, and Ad, resulting in the overexpression of a green fluorescent protein (eGFP) transgene marker. The transduction efficiency was assessed by flow cytometry, while cytotoxicity and apoptosis rate were detected by annexin V/propidium iodide (PI) stain. RESULTS: Ad had the highest transduction efficiency with 99% of the cells expressing the transgene, followed by EIAV (95%), HIV-1 (75%), and PFV (43%). However, the high transduction efficiency of Ad also resulted in the highest apoptosis rate (25%) in the corneal endothelial cells. There was no detectable difference in the toxicity between PFV and HIV-1 (10%). EIAV transduction had the lowest cytotoxicity, with only 3% of the cells being annexin V/PI positive. CONCLUSION: Compared to other vectors EIAV exhibited high transduction efficiency combined with low toxicity to corneal endothelial cells. Therefore, it is a powerful tool for gene therapy applications in selected corneal endothelial diseases.
Authors: Peng H Tan; Sven C Beutelspacher; Shao-An Xue; Yao-He Wang; Peter Mitchell; James C McAlister; D Frank P Larkin; Myra O McClure; Hans J Stauss; Mary A Ritter; Giovanna Lombardi; Andrew J T George Journal: Blood Date: 2005-01-25 Impact factor: 22.113
Authors: N Fischer; M Heinkelein; D Lindemann; J Enssle; C Baum; E Werder; H Zentgraf; J G Müller; A Rethwilm Journal: J Virol Date: 1998-02 Impact factor: 5.103
Authors: Alison S Bienemann; Enca Martin-Rendon; Anna S Cosgrave; Colin P J Glover; Liang-Fong Wong; Susan M Kingsman; Kyriacos A Mitrophanous; Nicholas D Mazarakis; James B Uney Journal: Mol Ther Date: 2003-05 Impact factor: 11.454
Authors: B Seitz; E Baktanian; E M Gordon; W F Anderson; L LaBree; P J McDonnell Journal: Graefes Arch Clin Exp Ophthalmol Date: 1998-08 Impact factor: 3.117