Adenoviral-based vectors
The present invention provides replication competent adenoviral vectors capable of expressing antigens from infectious pathogens, such as influenza virus. The adenoviral vectors can be used to vaccinate subjects against the infectious pathogens. The adenoviral vectors comprise heterologous sequences encoding the antigens. The heterologous sequences can be inserted into various locations in the adenoviral vectors, including in or near specific E3 deletions and/or integrated into the adenoviral hexon coding region. The adenoviral vectors can be derived from any adenoviral serotype, particularly an Ad4 or Ad7 serotype.
1. A vaccine comprising an adenoviral vector comprising a first heterologous sequence, wherein said adenoviral vector is replication competent and has a partial E3 deletion, and wherein the first heterologous sequence is integrated into a location containing the partial E3 deletion and is operably linked to a native adenovirus splice acceptor site and is under the control of the endogenous Major Late Promoter.
2. The vaccine of claim 1 , wherein the adenoviral vector is derived from Ad2, Ad3, Ad4, Ad5, Ad6, Ad7, Ad11, Ad20, Ad21, Ad22, Ad23, Ad24, Ad25, Ad26, Ad28, Ad34, Ad35, Ad40, Ad41, Ad48, Ad49, or Ad50.
3. The vaccine of claim 1 , wherein the partial E3 deletion comprises deletion of at least 1, 2 or 3 open reading frames within the E3 region.
4. The vaccine of claim 1 , wherein the partial E3 deletion does not alter the function of the E3 region.
5. The vaccine of claim 1 , wherein the partial E3 deletion comprises deletion of a region corresponding to ADP region of Ad5.
6. The vaccine of claim 1 , wherein the adenoviral vector is derived from Ad4 and the partial E3 deletion comprises a deletion of E3 24.8 k, E3 6.3 k, and E3 29.7 k.
7. The vaccine of claim 1 , wherein the adenoviral vector is derived from Ad7 and the partial E3 deletion comprises a deletion of E3 20.1 k, E3 20.6 k, and E3 7.7 k.
8. The vaccine of claim 1 , wherein the expression of the first heterologous sequence is under the control of the endogenous Major Late Promoter and tripartite leader sequence.
9. The vaccine of claim 1 , wherein the first heterologous sequence is operably linked to a native E3 24.8 k splice acceptor.
10. The vaccine of claim 1 , wherein the first heterologous sequence is operably linked to an adenoviral polyA signal sequence.
11. The vaccine of claim 10 , wherein the first heterologous sequence is operably linked to an Ad5 E3 polyA signal sequence.
12. The vaccine of claim 1 , wherein the first heterologous sequence encodes an immunogenic protein of an infectious pathogen.
13. The vaccine of claim 12 , wherein the infectious pathogen is selected from the group consisting of a virus, a bacterium, a protist, and a fungus.
14. The vaccine of claim 13 , wherein the infectious pathogen is influenza, human immunodeficiency virus, or human papilloma virus.
15. The vaccine of claim 13 , wherein the infectious pathogen is Bacillus, Shigella, Mycobacterium , or Plasmodium.
16. The vaccine of claim 1 , wherein the first heterologous sequence encodes influenza hemaglutinin, influenza neuraminidase, influenza M2, a multimer of M2e, a multimer of HTL epitopes, or a multimer of CTL epitopes.
17. The vaccine of claim 1 , wherein the first heterologous sequence comprises a first ORF encoding an immunogenic protein of an infectious pathogen and a second ORF encoding a multimer of epitopes from said infectious pathogen.
18. A vaccine comprising an adenoviral vector comprising a first heterologous sequence, wherein said adenoviral vector is derived from Ad2, Ad3, Ad4, Ad5, Ad6, Ad7, Ad11, Ad20, Ad21, Ad22, Ad23, Ad24, Ad25, Ad26, Ad28, Ad34, Ad35, Ad40, Ad41, Ad48, Ad49, Ad50, Ad C1, Ad C3, Ad C6, Ad C7, or Ad68, is replication competent, and has a full E3 deletion, and wherein the first heterologous sequence is operably linked to a native adenovirus splice acceptor site under the control of the endogenous Major Late Promoter.
19. A vaccine comprising an adenoviral vector comprising a first heterologous sequence and a second heterologous sequence, wherein the second heterologous sequence is integrated into an adenoviral hexon region, wherein the first heterologous sequence is integrated into an adenoviral non-hexon region, and wherein the adenoviral vector is replication competent, and wherein the first heterologous sequence is operably linked to a native adenovirus splice acceptor site under the control of the endogenous Major Late Promoter.
20. The vaccine of claim 19 , wherein the adenoviral vector is derived from Ad2, Ad3, Ad4, Ad5, Ad6, Ad7, Ad11, Ad20, Ad21, Ad22, Ad23, Ad24, Ad25, Ad26, Ad28, Ad34, Ad35, Ad40, Ad41, Ad48, Ad49, or Ad50.
21. The vaccine of claim 19 , wherein the adenoviral vector comprises a partial E3 deletion and wherein the first heterologous sequence is integrated into a location containing the partial E3 deletion.
22. The vaccine of claim 19 , wherein the second heterologous sequence is integrated into HVR1, HVR2, HVR4, or HVR5.
23. The vaccine of claim 19 , wherein the second heterologous sequence encodes a region of a membrane protein of a virus.
24. The vaccine of claim 19 , wherein the second heterologous sequence encodes an extracellular part of a conserved virus membrane protein.
25. The vaccine of claim 19 , wherein the second heterologous sequence encodes a region of an influenza M2 protein, an influenza Matrix CTL, an influenza NP epitope, one or more HVRs from an adenovirus of another serotype, or a combination thereof.
26. The vaccine of claim 19 , wherein the second heterologous sequence encodes one or more copies of M2e of influenza M2.
27. The vaccine of claim 19 , wherein the second heterologous sequence encodes one or more copies of M2e of influenza M2, wherein each M2 copy is integrated into a different HVR, and wherein the one or more copies of M2e are integrated into HVR1, HVR2, HVR4, HVR5 or a combination thereof.
28. The vaccine of claim 19 , wherein the second heterologous sequence comprises the sequence of SEQ ID NO. 318 (H5 M2e), SEQ ID NO. 321 (H7 M2e), SEQ ID NO. 327 (H9 M2e), SEQ ID NO. 312 (Human M2e), SEQ ID NO. 337 (NP), or SEQ ID NO. 336 (Matrix CTL).
29. The vaccine of claim 1 , 18 , or 19 , which is formulated for oral, intranasal, sublingual, intravesical, rectal, or intravaginal administration.
30. A vaccine of claim 1 , 18 , or 19 , further comprising an acceptable carrier.
31. A dosage unit of the vaccine of claim 1 , 18 , or 19 , wherein a single dose comprises about 10 3 to about 10 13 adenoviral particles.
32. A method of inducing an immune response to an infectious pathogen in a subject comprising administering to the subject the vaccine of claim 1 , 18 , or 19 .
33. The method of claim 32 , wherein one or more doses of the vaccine is administered to the subject.
34. The method of claim 32 , wherein the infectious pathogen is influenza, HIV, HPV, Bacillus, Plasmodium, Mycobacteria , or Shigella.
35. The method of claim 32 , wherein the subject has an infection induced by said infectious pathogen.