Title
Electroporating human corneal epithelial cells with interleukin 10 and Fas ligand pDNA Electroporating human corneal epithelial cells with interleukin 10 and Fas ligand pDNA
Author
Faculty/Department
Faculty of Medicine and Health Sciences
Publication type
article
Publication
Singapore ,
Subject
Human medicine
Source (journal)
Asia-Pacific journal of ophthalmology / National University Hospital of Singapore. Department of Ophthalmology. - Singapore
Volume/pages
3(2014) :1 , p. 56-63
ISSN
0129-1653
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Purpose: To develop an optimal electroporation protocol for plasmid (pDNA) transfection of a human corneal epithelial cell (hCEC) line and investigate the immunomodulatory capacity of interleukin 10 and Fas ligand (FasL) transfection on hCECs. Design: A controlled experimental study. Methods: Human corneal epithelial cells were electroporated with pDNA encoding enhanced green fluorescent protein, interleukin 10, or FasL. Supernatants were analyzed for cytokine secretion using enzyme-linked immunosorbent assay. To test potential immunosuppression, electroporated hCECs were cocultured with allogeneic peripheral blood mononuclear cells, and the supernatants analyzed for interferon γ production. Results: Maximum transfection efficiencies were obtained using optimized settings, and transgene expression was detected up to 13 days following transfection. Interleukin 10 levels peaked at day 4 and FasL at day 2 following electroporation. Coculture supernatants showed significantly lower levels of interferon γ in the modulated groups compared with control. Conclusions: Our results demonstrate highly efficient transfection of hCECs using an optimized electroporation protocol. Interleukin 10 and FasL may provide a means of immune modulation of corneal epithelial cells.
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