|
Title
|
|
|
|
Electroporating human corneal epithelial cells with interleukin 10 and Fas ligand pDNA
| |
|
Author
|
|
|
|
| |
|
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. |
| |
|
Language
|
|
|
|
English
| |
|
Source (journal)
|
|
|
|
Asia-Pacific journal of ophthalmology / National University Hospital of Singapore. Department of Ophthalmology. - Singapore
| |
|
Publication
|
|
|
|
Singapore
:
2014
| |
|
ISSN
|
|
|
|
0129-1653
| |
|
DOI
|
|
|
|
10.1097/APO.0000000000000034
| |
|
Volume/pages
|
|
|
|
3
:1
(2014)
, p. 56-63
| |
|
Full text (Publisher's DOI)
|
|
|
|
| |
|