Defective adenoviruses and corresponding complementation lines
Novel defective adenoviruses for the transfer and expression of an exogenous nucleotide sequence in a host cell or organism. The invention also relates to novel complementation lines and to the process for the preparation of these novel defective adenoviruses and their use in therapy and to a pharmaceutical composition containing same.
1. A stock of recombinant adenoviral vector produced in a complementation cell line, wherein said recombinant adenoviral vector is deficient in one or more essential gene functions of the E1 region of the adenoviral genome, and wherein there is an elimination of the adenoviral sequences which are in common to said adenoviral genome and adenoviral E1 sequences integrated in said complementation cell line to avoid a recombination event sufficient to generate a non-defective recombinant adenovirus.
2. The stock of claim 1 , wherein said recombinant adenoviral vector is deficient in two essential gene functions.
3. The stock of claim 2 , wherein said recombinant adenoviral vector is deficient in one or more essential gene functions of each of the E1 and E2 regions of the adenoviral genome.
4. The stock of claim 2 , wherein said recombinant adenoviral vector is deficient in one or more essential gene functions of each of the E1 and E4 regions of the adenoviral genome.
5. The stock of claim 1 , wherein said recombinant adenoviral vector is deficient in three essential gene functions.
6. The stock of claim 5 , wherein said recombinant adenoviral vector is deficient in one or more essential gene functions of each of the E1, E2 and E4 regions of the adenoviral genome.
7. A system comprising (i) a cell having a cellular genome comprising at least one adenoviral sequence, which is operably linked to a promoter, and which complements in trans for a deficiency in one or more essential gene functions of the E1 region of an adenoviral genome, and (ii) an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene functions of the E1 region of said adenoviral genome, wherein there is an elimination of the adenoviral sequences which are in common to said adenoviral genome and the at least one adenoviral sequence in said cell to avoid a recombination event sufficient to generate a non-defective adenovirus.
8. A method of propagating an adenoviral vector that is deficient in one or more essential gene functions of one or more regions of an adenoviral genome, which method comprises propagating an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene functions of the E1 region in a cell having a cellular genome comprising at least one DNA sequence that complements in trans said deficiency, wherein there is an elimination of the adenoviral sequences which are in common to said adenoviral genome and adenoviral E1 sequences integrated in said cell to avoid a recombination event resulting in a non-defective adenoviral vector.