TRANS COMPLEMENTATION OF AN E1A-DELETED ADENOVIRUS WITH CODELIVERED E1A SEQUENCES TO MAKE RECOMBINANT ADENOVIRAL PRODUCER CELLS

Replication-incompetent adenovirus is conventionally produced by cells that supply replication-enabling proteins from viral sequences presen t in trans. As an alternative means of recombinant adenovirus producti on, replication-enabling E1A sequences were cotransduced into human pr ostate carcinoma cells infected with an E1A-deleted adenovirus contain ing a luciferase expression cassette. The replication-enabling plasmid was cotransduced by ionic linkage to the recombinant adenovirus exter ior. Cells cotransduced with the replication-enabling plasmid made new adenovirus with titers up to 8 x 10(6) in the supernatants 72-120 hr after transduction. Like the parent virus, the virus present in the co transduced supernatants and lysates was capable of transferring lucife rase activity to new cells. The virus present in the cotransduced cell supernatants was amplified and shown to be identical to the parent vi rus by genomic analysis. It was concluded that simultaneous addition o f a replication-defective adenovirus and a replication-enabling gene s equence in a trans configuration converts some of the cotransduced cel ls into recombinant adenovirus-producing cells.