Recombinant poxvirus vector comprising tetanus toxin fragment C
The present invention relates to a recombinant poxvirus comprising tetanus toxin fragment C for improved immunogenicity of an antigen and related methods and uses. Specifically, the present invention generally relates to genetically engineered (recombinant) poxvirus vectors comprising a tetanus toxin fragment C (TTC) coding sequence operably linked to a bacterial antigenic determinant as well as to uses thereof, e.g., to affect an immune response in a subject.
1. A recombinant poxvirus comprising a fusion protein coding sequence, the fusion protein comprising a tetanus toxin fragment C (TTC) and a Bacillus anthracis Protective antigen (PA) protein, wherein the coding sequence for the TTC encodes for amino acids 761-1213 of SEQ ID NO:2, and wherein the fusion protein enhances an immune response against Bacillus anthracis as compared with an immune response by a recombinant poxvirus comprising PA alone.
2. The recombinant poxvirus according to claim 1 , wherein the TTC coding sequence comprises nucleotides 2281-3642 of SEQ ID NO: 1.
3. The recombinant poxvirus according to claim 1 , wherein in the TTC is fused to the PA protein via a linker sequence.
4. The recombinant poxvirus of claim 1 , wherein the poxvirus is a vaccinia virus.
5. The recombinant poxvirus of claim 4 , wherein the vaccinia virus is a Modified Vaccinia Virus Ankara (MVA).
6. The recombinant poxvirus of claim 5 , wherein the MVA is MVA-BN.
7. The recombinant poxvirus of claim 1 , wherein the recombinant poxvirus further comprises a coding sequence for at least one Bacillus anthracis antigenic determinant selected from the group consisting of lethal factor (LF) and edema factor (EF).
8. The recombinant poxvirus according to claim 1 , further comprising a nucleic acid encoding a co-stimulatory molecule.
9. The recombinant poxvirus of claim 8 , wherein the co-stimulatory molecule is CD70, IL-12 and/or IL-15.
10. A vaccine or medicament for treating and/or preventing anthrax comprising the recombinant poxvirus of claim 6 .
11. An isolated cell comprising the recombinant poxvirus according to claim 6 .
12. A kit comprising the recombinant poxvirus according to claim 6 in a first vial or container for a first administration and in a second vial or container for a second administration.
13. A method for preventing or treating anthrax in a subject, the method comprising administering to the subject a recombinant poxvirus comprising a coding sequence for a fusion protein, the fusion protein comprising a tetanus toxin fragment C (TTC) and a Bacillus anthracis Protective antigen (PA) protein, wherein the coding sequence for the TTC encodes for amino acids 761-1213 of SEQ ID NO:2, and wherein said fusion protein enhances an immune response against Bacillus anthracis as compared with an immune response by a recombinant poxvirus comprising PA alone.
14. The method according to claim 13 , wherein the TTC coding sequence comprises nucleotides 2281-3642 of SEQ ID NO: 1.
15. The method according to claim 13 , wherein in the TTC is fused to the PA protein via a linker sequence.
16. The method of claim 13 , wherein the poxvirus is a vaccinia virus.
17. The method of claim 16 , wherein the vaccinia virus is a Modified Vaccinia Virus Ankara (MVA).
18. The method of claim 17 , wherein the MVA is MVA-BN.
19. A method of generating a recombinant poxvirus comprising a coding sequence for a fusion protein, the fusion protein comprising a tetanus toxin fragment C (TTC) and a Bacillus anthracis Protective antigen (PA) protein, wherein the coding sequence for the TTC encodes for amino acids 761-1213 of SEQ ID NO:2, and wherein said fusion protein enhances an immune response against Bacillus anthracis as compared with an immune response by a recombinant poxvirus comprising PA alone the method comprising:
a) infecting a host cell with a poxvirus;
b) transfecting the infected cell with a recombinant vector comprising the fusion protein, said vector further comprising a genomic poxvirus sequence capable of directing the integration of the TTC and antigenic determinant coding sequences into the poxvirus genome; and
c) identifying, isolating and, optionally, purifying the generated recombinant poxvirus.