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[Comparison of several viral vectors for gene therapy of corneal endothelial cells].
- Source :
-
Der Ophthalmologe : Zeitschrift der Deutschen Ophthalmologischen Gesellschaft [Ophthalmologe] 2005 Dec; Vol. 102 (12), pp. 1168-74. - Publication Year :
- 2005
-
Abstract
- 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.<br />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.<br />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.<br />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.
- Subjects :
- Animals
Cell Line
Corneal Diseases genetics
Corneal Diseases therapy
DNA, Viral administration & dosage
DNA, Viral genetics
Gene Expression Profiling
Genetic Therapy methods
Genetic Vectors genetics
Green Fluorescent Proteins genetics
Horses
Humans
Mice
Adenoviridae genetics
Epithelium, Corneal metabolism
Green Fluorescent Proteins metabolism
HIV-1 genetics
Infectious Anemia Virus, Equine genetics
Spumavirus genetics
Transfection methods
Subjects
Details
- Language :
- German
- ISSN :
- 0941-293X
- Volume :
- 102
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Der Ophthalmologe : Zeitschrift der Deutschen Ophthalmologischen Gesellschaft
- Publication Type :
- Academic Journal
- Accession number :
- 15886987
- Full Text :
- https://doi.org/10.1007/s00347-005-1230-6