The impact of developmental stage, route of administration and the immune system on adenovirus-mediated gene transfer.

Important aspects of successful adenovirus gene transfer include the amount and persistence of gene expression, the ability to readminister virus and the localization of virus-directed gene expression to target organs. Our objective in this study was to use a single recombinant adenovirus bearing a quantifiable reporter gene [chloramphenicol acetyltransferase (CAT)] to establish the parameters which define the limits of adenovirus gene expression in a rat model. First, we determined how the route of virus administration affected the amount, duration and distribution of expression in different tissues and in rats of different developmental stages. All routes resulted in infection of all tissues tested. Surprisingly, the most efficient and widespread gene transfer was achieved by intracardiac muscle injection. The high levels of CAT protein that can be produced in a liver (< or = 1.7 mg) or a heart (< or = 196 micrograms) 5 days after infection suggest that the amount of gene product will not be a limitation in the use of adenovirus. Following peak activity at 5 days after infection, a gradual decline of CAT expression was observed in all tissues assayed; by 80 days neither CAT activity nor adenovirus DNA were detectable. In addition, adults could not be boosted by a second administration of virus, presumably due to the presence of high levels of neutralizing antibodies. The limited persistence of gene expression could be circumvented when virus was injected into neonates. Blocking T lymphocyte expansion by cyclosporine enhanced the persistence of CAT gene product over a 25-day period in heart and lung but not in liver compared with control animals.(ABSTRACT TRUNCATED AT 250 WORDS)