title: An experimental study on in situ transfection of the interleukin-1 receptor antagonist (IL-1ra) gene into the rat cornea.
abstract: OBJECTIVE
  To investigate the efficiency of the transfection of PEGFP-IL-1ra plasmid via cation polymer mediation (poly-ethylenimine, PEI) by injection into the corneal stroma.
  
  
  METHODS
  Plasmid PEGFP-hIL-1ra recombinants were constructed and transferred into corneal endothelial cells via cation polymer mediation. Plasmid PEGFP-hIL-1ra recombinants and/or PEI-in-vivo were injected into the corneal stroma of Wistar rats. Corneas were harvested at different time points (days 3, 6, 14 and 21) after injection. The expression of IL-1ra after transfection was studied by fluorescence microscopy, transmission electron microscopy, real time polymerase chain reaction (RT-PCR) and immunohistochemistry.
  
  
  RESULTS
  Plasmid PEGFP-hIL-1ra recombinants were constructed successfully. After injection of pEGFP-hIL-1ra plasmid into the cornea, IL-1ra mRNA expression was detected in the corneal stroma and reached a peak on day 6. IL-1ra-GFP granules could be observed in every layer of the cornea in the PEGFP-hIL-1ra recombinant group by transmission electron microscopy, but not in the negative control (PEI-in-vivo) group. P63 immunocytochemical staining in the corneal epithelium was positive in both groups. There was no impairment in the ultrastructure of cells in both groups.
  
  
  CONCLUSIONS
  By direct injection of PEGFP-hIL-1ra into the corneal stroma and mediation by the cation polymer, the IL-1ra gene could be transferred and expressed in corneal tissue efficiently. This may be a novel technique for gene transfection to the cornea in situ.
authors: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou, PR China.
authors: Jin
authors: J
authors: Yuan
authors: Yi
authors: Y
authors: Liu
authors: Huang
authors: H
authors: Weilan
authors: Shiyou
authors: S
authors: Zhou
authors: Jiaqi
authors: J
authors: Chen
mesh_terms: D000818:Animals; D003315:Cornea / Q000378:metabolism* / Q000648:ultrastructure; D003316:Corneal Diseases / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D004195:Disease Models, Animal; D004587:Electrophoresis, Agar Gel; D005260:Female; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D053590:Interleukin 1 Receptor Antagonist Protein / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics*; D008297:Male; D017348:Microscopy, Electron, Scanning Transmission; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*
chemical_list: D053590:Interleukin 1 Receptor Antagonist Protein; D012333:RNA, Messenger; D011994:Recombinant Proteins