Recombinant adenovirus vector having a reduced side effect
The present invention provides a novel adenovirus vector for which inflammation during the in vivo administration thereof is alleviated by inhibiting the induction of expression of an adenovirus gene by a foreign promoter inserted into the adenovirus genome, and a method for producing the vector, a cell line for use in the production of the recombinant adenovirus vector, or a gene therapy method using the recombinant adenovirus vector.
1. A recombinant adenovirus vector, for which inflammation during in vivo administration thereof is alleviated as compared with a first generation adenovirus vector lacking the E1 gene, said recombinant adenovirus vector having the following characteristics:
(a) deletion of the adenovirus E1A and E1B genes;
(b) retention of at least ORF3 of the adenovirus E4 gene;
(c) presence of a foreign gene under control of a foreign promoter;
(d) relocation of the adenovirus protein IX gene to a position in said vector at which expression of said protein IX gene is not induced by said foreign promoter, said position being at least 18 kb or more from the foreign promoter, and either: (i) between the adenovirus L3 gene and the adenovirus E2A gene, or (ii) between the E4 gene and the 3′ ITR:
wherein said vector encodes a virus particle having infectivity and thermal stability equivalent to that of an adenovirus wild-type strain.
2. The recombinant adenovirus vector according to claim 1 , wherein the foreign gene under the control of the foreign promoter has been inserted into the E1A and E1B gene deletion site.
3. The recombinant adenovirus vector according to claim 1 , further comprising a deletion of all or part of an additional adenovirus gene.
4. The recombinant adenovirus vector according to claim 1 , further comprising a deletion of all or part of the adenovirus E3 gene.
5. The recombinant adenovirus vector according to claim 1 , further comprising a deletion of all or part of the adenovirus E2A gene.
6. The recombinant adenovirus vector according to claim 1 , wherein in characteristic (b) only ORF3 of the E4 gene has been retained.
7. The recombinant adenovirus vector according to claim 1 , wherein the protein IX gene has been relocated to a position between the adenovirus L3 gene and the adenovirus E2A gene.
8. The recombinant adenovirus according to claim 1 , wherein the protein IX gene has been relocated to a position between the E4 gene and the 3′′ ITR.
9. The recombinant adenovirus vector according to claim 1 , wherein the foreign promoter contains an element derived from an animal virus.
10. The recombinant adenovirus vector according to claim 9 , wherein the foreign promoter contains a cytomegalovirus (CMV) enhancer.
11. The recombinant adenovirus vector according to claim 10 , wherein the foreign promoter is a CMV promoter.
12. The recombinant adenovirus vector according to claim 10 , wherein the foreign promoter is a CAG promoter.
13. The recombinant adenovirus vector according to claim 9 , wherein the foreign promoter is a Rous sarcoma virus (RSV) promoter or a SRα promoter.
14. The recombinant adenovirus vector according to claim 1 , wherein said recombinant adenovirus vector is a human adenovirus.
15. The recombinant adenovirus vector according to claim 14 , wherein said recombinant adenovirus vector is a type 2 adenovirus or a type 5 adenovirus.
16. The recombinant adenovirus vector according to claim 2 , wherein said recombinant adenovirus vector comprises an additional deletion of all or part of an adenovirus gene.
17. The recombinant adenovirus vector according to claim 9 , wherein said animal virus is a mammalian virus.