[Comparison of several viral vectors for gene therapy of corneal endothelial cells].
OBJECTIVE
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.


METHODS
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.


CONCLUSIONS
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.
Beutelspacher
S C
SC
Augenklinik, Ruprecht-Karls-Universität Heidelberg. beutelspacher@urz.uni-heidelberg.de
Serbecic
N
N
Tan
P
P
McClure
M O
MO
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D003316:Corneal Diseases / Q000235:genetics / Q000628:therapy; D004279:DNA, Viral / Q000008:administration & dosage / Q000235:genetics; D019573:Epithelium, Corneal / Q000378:metabolism*; D020869:Gene Expression Profiling; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics*; D006736:Horses; D006801:Humans; D004860:Infectious Anemia Virus, Equine / Q000235:genetics*; D051379:Mice; D016092:Spumavirus / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D004279:DNA, Viral; D049452:Green Fluorescent Proteins