Adenovirus gene therapy for hypercholesterolemia, thrombosis and restenosis.

Human adenoviruses are non-enveloped, icosahedral viruses, approximately 130 nm in diameter. The linear, double-stranded DNA viral genome containing covalently attached terminal protein derived from a unique replication process is almost 36 Kb in length. Although forty nine serotypes of human adenovirus in six serological subgroups have been identified, only serotypes 2 and 5 of subgroup C have been extensively employed as gene transfer vectors. Recombinant helper-independent adenovirus vectors are easily constructed, analyzed and propagated using standard Ž recombinant DNA and virological techniques reviewed in w x. 1,2 . Foreign genes can be inserted into the adenovirus genome in a variety of locations to generate recombinant Ž . vectors, although substitution of early region 1 E1 has been most widely used. Replacement of E1, since it is required for the efficient expression of the remainder of the viral genome, generates a vector which can only be propagated in a complementing cell line which supply the missing E1A functions in trans from an integrated copy of the appropriate fragment of the viral genome. Such E1 w x replacement vectors are typically propagated in the 293 3 w x or 911 4 cell lines. Recombinant viruses produced in this manner can be stably propagated to a titer of 10–10 plaque forming units per ml and are readily purified free of