EXPRESSION OF INTERLEUKIN-4 IN CULTURED CORNEAS FOLLOWING GENTRANSFER
U. Pleyer1, P. Rieck1, C. Hartmann1, H.-D. Volk2, T. Ritter2
The transfer of recombinant DNA into specific organs may offer new therapeutic options in transplantation medicine. Ex vivo gene therapy seems especially attractive to modulate the allogeneic immune response in keratoplasty. This study examined the potential to express the Th2-cytokine Interleukin-4 (IL-4) in organ cultured corneas.
Methods: Wistar rat corneas were transfected with the recombinant adenovirus El/E3 containing the AdrlL-4 gene. Ex vivo infections were performed using two different concentrations of AdrlL-4 (2x107pfu and 2x108 pfu) in DMEM/2% FCS for 3 hours. The infection medium was replaced by DMEM/10%FCS for organ culture. After 24, 72 and 144 hours aliquots of supernatants were assayed for IL-4 by ELISA. Control corneas were infected with a reporter construct (Adb-Gal).
Results: The infection of rat corneas with the adenoviral vector AdlL-4 in vitro resulted in high expression of IL-4 after 72 hours in a dose dependent manner (2x107 pfu, conc: 230pg IL-4/ml; 2x 108 pfu, conc: > 30ng IL-4/ml). Production of IL-4 was already detectable after 24 hours for the corneas infected with the higher concentration of AdrlL-4. The infection of rat corneas with Adb- Gal did not lead to any IL-4 production in vitro.
Conclusions: In this study we have shown the successful adenoviral-mediated gene transfer of the Th2-cytokine IL-4 in rat cornea in vitro. Further studies are in progress to investigate the role of IL-4 in the prevention of an allogeneic immune response against genetically modified cornea transplants.
Supported by DFG (Pl 150/9-1 and Vo 489/6-1)
1Dept. of Ophthalmology, Charité, Campus Virchow Klinikum, 13353 Berlin
2Institute of Medical Immunology, Charité, Berlin, Germany