title: Electroporating Human Corneal Epithelial Cells With Interleukin 10 and Fas Ligand pDNA.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
authors.affiliation: From the *Department of Ophthalmology and †Center for Cell Therapy and Regenerative Medicine, Antwerp University Hospital; and ‡Laboratory of Experimental Haematology, Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Edegem, Belgium.
authors.forename: Nadia
authors.initials: N
authors.lastname: Zakaria
authors.forename: Nathalie
authors.initials: N
authors.lastname: Cools
authors.forename: Zwi
authors.initials: Z
authors.lastname: Berneman
authors.forename: Marie-Jose
authors.initials: MJ
authors.lastname: Tassignon