Vergleich verschiedener viraler Vektoren zur Gentherapie von Hornhautendothelzellen

Abstract

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. 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. 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. 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.