TNF receptor secretion after ex vivo adenoviral gene transfer to cornea and effect on in vivo graft survival.
OBJECTIVE
To explore the potential for adenovirus-mediated ex vivo gene transfer of a soluble tumor necrosis factor (TNF) receptor and evaluate the effect of transplanting the adenovirally transplanted corneas in vivo.


METHODS
Rabbit corneal segments were transfected with replication-deficient adenovirus (AdTNFR) encoding a soluble TNF receptor fusion protein (TNFR-Ig). Production of TNFR-Ig was measured by using ELISA and bioassay. Corneas were transfected ex vivo with AdTNFR and then transplanted in vivo. Survival of AdTNFR-transfected corneas was compared with that of those treated either with a null vector control adenovirus (Ad0) or nontransfected control corneas.


RESULTS
Ex vivo production of a molecule with TNF blocking bioactivity from AdTNFR-transfected corneas was demonstrated over a period of 4 weeks. Transplanted AdTNFR-transfected corneas showed a marginally increased survival time in vivo over nontransfected control corneas, but a significantly increased survival time over Ad0-treated control corneas. Ad0 treatment of corneal allografts before transplantation had a proinflammatory effect and accelerated the onset of corneal endothelial rejection.


CONCLUSIONS
Adenoviral gene transfer is an effective means of transferring a gene encoding soluble TNFR-Ig to corneal endothelium, and ex vivo production of a biologically active secreted molecule was demonstrated for 4 weeks. However, in vivo, only a marginally increased survival was seen compared with control corneas. The introduction of this transgene using a less immunogenic vector may demonstrate prolongation of corneal allograft survival.
Rayner
S A
SA
Department of Immunology, Division of Medicine, Imperial College School of Medicine, Hammersmith Hospital, London, UK.
Larkin
D F
DF
George
A J
AJ
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016039:Corneal Transplantation / Q000276:immunology*; D003673:Defective Viruses; D004728:Endothelium, Corneal / Q000378:metabolism* / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006085:Graft Survival / Q000276:immunology; D007074:Immunoglobulin G / Q000502:physiology*; D011817:Rabbits; D018124:Receptors, Tumor Necrosis Factor / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000502:physiology; D014162:Transfection; D014184:Transplantation, Homologous
D007074:Immunoglobulin G; D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins