Avipox recombinants expressing foot and mouth disease virus genes
View Patent ↗The present invention relates to modified poxviral vectors and to methods of making and using the same. In particular, the invention relates to recombinant avipox that expresses gene products of foot and mouth disease virus (FMDV), and to compositions or vaccines that elicit immune responses directed to FMDV gene products and which can confer protective immunity against infection by FMDV.
1. A recombinant avipox vector selected from canarypox or fowlpox, comprising at least one nucleic acid molecule encoding one or more foot-and-mouth disease virus (FMDV) antigen(s), wherein the nucleic acid molecule is inserted into a single insertion site and wherein:
(a) the nucleic acid molecule is operably linked to an early or early late promoter sequence; or
(b) the nucleic acid molecule is operably linked to a relatively weak promoter;
wherein if the avipox vector is canarypox, the promoter is in the opposite orientation to the flanking sequences and wherein expression levels of the FMDV antigen(s) are decreased compared with expression levels of the FMDV antigen(s) under a relatively strong promoter.
2. The avipox vector of claim 1 , wherein the antigen is selected from the group consisting of FMDV VP1, VP2, VP3, VP4, 2A, 2B, and 3C.
3. The avipox vector of claim 1 , wherein the nucleic acid molecule encoding one or more foot-and-mouth disease virus (FMDV) antigen(s) is a cDNA encoding FMDV P1 region and a cDNA encoding FMDV 3C protease.
4. The avipox vector of claim 1 , wherein the promoter sequence is selected from the group consisting of H6 vaccinia promoter, 13L vaccinia promoter, 42K poxviral promoter, 7.5K vaccinia promoter and Pi vaccinia promoter.
5. The avipox vector of claim 1 , wherein the promoter is the H6 vaccinia promoter, which is mutated such that expression levels of the FMDV antigen(s) are decreased compared with expression levels of the FMDV antigen(s) under a wild type H6 vaccinia promoter.
6. The avipox vector of claim 1 , wherein the vector comprises a C6 insertion locus, and wherein flanking sequences of the C6 insertion locus promote homologous recombination of the FMDV antigens with the C6 insertion locus.
7. The avipox vector of claim 6 , wherein the flanking sequences comprise C6L and C6R open reading frames of avipox.
8. The avipox vector of claim 1 , wherein the vector comprises a F8 insertion locus, and wherein flanking sequences of the F8 insertion locus promote homologous recombination of the FMDV antigens with the F8 insertion locus.
9. The avipox vector of claim 8 , wherein the flanking sequences comprise F8L and F8R open reading frames of avipox.
10. A recombinant avipox virus selected from ALVAC or fowlpox, comprising at least one nucleic acid molecule encoding one or more foot-and-mouth disease virus (FMDV) antigen(s)), wherein the nucleic acid molecule is inserted into a single insertion site and wherein:
(a) the nucleic acid molecule is operably linked to an early or early late promoter sequence; or
(b) the nucleic acid molecule is operably linked to a relatively weak promoter;
wherein if the avipox vector is ALVAC, the promoter is in the opposite orientation to the flanking sequences and wherein expression levels of the FMDV antigen(s) are decreased compared with expression levels of the FMDV antigen(s) under a relatively strong promoter.
11. The avipox virus of claim 10 , wherein the antigen is selected from the group consisting of FMDV VP1, VP2, VP3, VP4, 2A, 2B, and 3C.
12. The avipox virus of claim 10 , wherein the nucleic acid molecule encoding one or more foot-and-mouth disease virus (FMDV) antigen(s) is a cDNA encoding FMDV P1 region and a cDNA encoding FMDV 3C protease.
13. The avipox virus of claim 10 , wherein the promoter sequence is selected from the group consisting of H6 vaccinia promoter, 13L vaccinia promoter, 42K poxviral promoter, 7.5K vaccinia promoter and Pi vaccinia promoter.
14. The avipox virus of claim 10 , wherein the promoter is the H6 vaccinia promoter, which is mutated such that expression levels of the FMDV antigen(s) are decreased compared with expression levels of the FMDV antigen(s) under a wild type H6 vaccinia promoter.
15. The avipox virus of claim 10 , wherein the vector comprises a C6 insertion locus, and wherein flanking sequences of the C6 insertion locus promote homologous recombination of the FMDV antigens with the C6 insertion locus.
16. The avipox vector of claim 15 , wherein the flanking sequences comprise C6L and C6R open reading frames of avipox.
17. The avipox virus of claim 10 , wherein the vector comprises a F8 insertion locus, and wherein flanking sequences of the F8 insertion locus promote homologous recombination of the FMDV antigens with the F8 insertion locus.
18. The avipox vector of claim 17 , wherein the flanking sequences comprise F8L and F8R open reading frames of avipox.
19. A recombinant avipox virus, wherein the avipox virus is selected from the group consisting of vCP2186, vCP2181, vCP2176, and vFP2215.
20. A method of eliciting an immune response to FMDV in a subject, comprising administering the vector of claim 1 to the subject.
21. A method of eliciting an immune response to FMDV in a subject, comprising administering the virus of claim 10 to the subject.
22. A method of eliciting an immune response to FMDV in a subject, comprising administering the virus of claim 19 to the subject.