Ex vivo adenovirus-mediated gene transfer and immunomodulatory protein production in human cornea.

One attractive strategy to prevent or control allograft rejec- transcription after infection. In contrast, adenoviral DNA tion is to genetically modify the donor tissue before trans- persisted for at least 56 days. Subsequently, we examined plantation. In this study, we have examined the feasibility the expression of a potential therapeutic gene, CTLA-4 Ig of gene transfer to human corneal endothelium, using a fusion protein. Following infection of human corneas with number of recombinant adenovirus constructs. Ex vivo adenoviral vectors encoding CTLA-4 Ig protein, high levels infection of human corneas with adenoviral vectors con- of the fusion protein were detected in corneal culture taining lacZ, under transcriptional control of either cyto- supernatants for up to 28 days. This protein was funcmegalovirus (CMV) or Rous sarcoma virus (RSV) pro- tionally active, as determined by binding to B7.1 (CD80)moters, provided high-level gene expression, which was expressing transfectants. This study suggests that genetic largely restricted to endothelium. Expression of the alteration of donor cornea before transplantation is a feasreporter gene persisted at relatively high levels for up to 7 ible approach for preventing or controlling allograft rejecdays, followed by a decline to indetectable levels by 28 tion. Similar gene-based strategies might also be feasible days. RT-PCR analysis of lacZ transcription showed a to prevent rejection of other transplanted tissues or organs. similar picture with a short period (3‐7 days) of RNA