Sa. Rayner et al., TNF receptor secretion after ex vivo adenoviral gene transfer to cornea and effect on in vivo graft survival, INV OPHTH V, 42(7), 2001, pp. 1568-1573
PURPOSE. TO explore the potential for adenovirus-mediated ex vivo gene tran
sfer of a soluble tumor necrosis factor (TNF) receptor and evaluate the eff
ect of transplanting the adenovirally transplanted corneas in vivo.
METHODS. Rabbit corneal segments were transfected with replication-deficien
t adenovirus (AdTNFR) encoding a soluble TNF receptor fusion protein (TNFR-
Ig). Production of TNFR-Ig was measured by using ELISA and bioassay. Cornea
s were transfected ex vivo with AdTNFR and then transplanted in vivo. Survi
val of AdTNFR-transfected corneas was compared with that of those treated e
ither with a null vector control adenovirus (AdO) or nontransfected control
corneas.
RESULTS. EX vivo Production of a molecule with TNF blocking bioactivity fro
m AdTNFR-transfected corneas was demonstrated over a period of 4 weeks. Tra
nsplanted AdTNFR-transfected corneas showed a marginally increased survival
time in vivo over nontransfected control corneas, but a significantly incr
eased survival time over Ad0-treated control corneas. Ad0 treatment of corn
eal allografts before transplantation had a proinflammatory effect and acce
lerated 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 produc
tion of a biologically active secreted molecule was demonstrated for 4 week
s. However, in vivo, only a marginally increased survival was seen compared
with control corneas. The introduction of this transgene using a less immu
nogenic vector may demonstrate prolongation of corneal allograft survival.