Compositions and methods for generating an immune response to treat or prevent malaria
The compositions and methods are described for generating an immune response to a Plasmodium antigen. The compositions and methods described herein relate to a modified vaccinia Ankara (MVA) vector encoding one or more viral antigens for generating a protective immune response to malaria by expressing the Plasmodium antigen in the subject to which the MVA vector is administered. The compositions and methods of the present invention are useful both prophylactically and therapeutically and may be used to prevent and/or treat malaria.
1. A recombinant modified vaccinia Ankara (MVA) viral vector comprising:
i) a first nucleic acid sequence encoding a Plasmodium falciparum immunogenic polypeptide comprising a circumsporozoite protein (CSP) with 21 asparagine (N)-alanine (A)-asparagine (N)-proline (P) (NANP) repeats (CSP21R),
ii) a second nucleic acid sequence encoding a Plasmodium falciparum immunogenic polypeptide comprising a gametocyte surface protein P230 (Pfs230), the Plasmodium falciparum immunogenic Pfs230 polypeptide further comprising a transmembrane domain of a glycoprotein (GP) of Marburg virus, and
iii) a third nucleic acid encoding a Marburg virus VP40 matrix protein;
wherein the first nucleic acid, the second nucleic acid sequence, and the third nucleic acid sequence are each under the control of a promoter compatible with poxvirus expression systems.
2. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence, the second nucleic acid sequence, and the third nucleic acid sequence are inserted into the MVA in a natural deletion site, a modified natural deletion site, between essential MVA genes, or between non-essential MVA genes.
3. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence is inserted between MVA genes I8R and G1L or between MVA genes A50R and B1R.
4. The recombinant MVA viral vector of claim 1 , wherein the second nucleic acid sequence is inserted between MVA genes I8R and G1L or between MVA genes A50R and B1R.
5. The recombinant MVA viral vector of claim 1 , wherein the third nucleic acid sequence is inserted between MVA genes I8R and G1L or between MVA genes A50R and B1R.
6. The recombinant MVA viral vector of claim 1 , wherein each promoter is selected from the group consisting of a Pm2H5 promoter, a Psyn II promoter, and a mH5 promoter.
7. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence, the second nucleic acid sequence, and the third nucleic acid sequence are optimized by one or more methods selected from the group consisting of: changing selected codons to other synonymous codons that are optimal for protein expression by MVA, interrupting homopolymer stretches using silent mutations, and interrupting transcription terminator motifs using silent mutations.
8. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid comprises the nucleic acid sequence of SEQ ID NO: 1.
9. The recombinant MVA viral vector of claim 1 , wherein the Plasmodium falciparum CSP21R immunogenic polypeptide comprises the amino acid sequence of SEQ ID NO: 2, or an amino acid sequence at least 95% identical thereto.
10. The recombinant MVA viral vector of claim 1 , wherein the Plasmodium falciparum Pfs230 immunogenic polypeptide comprises the amino acid sequence of SEQ ID NO: 6, or an amino acid sequence at least 95% identical thereto, further comprising a transmembrane domain of a glycoprotein (GP) of Marburg virus.
11. The recombinant MVA viral vector of claim 1 , wherein the third nucleic acid comprises the nucleic acid sequence of SEQ ID NO: 7.
12. The recombinant MVA viral vector of claim 1 , wherein the Marburg virus VP40 matrix protein comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence at least 95% identical thereto.
13. A recombinant modified vaccinia Ankara (MVA) viral vector comprising:
i) a first nucleic acid encoding a Plasmodium falciparum CSP21R immunogenic polypeptide comprising the amino acid sequence of SEQ ID NO: 2, or an amino acid sequence at least 95% identical thereto,
ii) a second nucleic acid sequence encoding a Plasmodium falciparum Pfs230 immunogenic polypeptide comprising the amino acid sequence of SEQ ID NO: 6, or an amino acid sequence at least 95% identical thereto, further comprising a transmembrane domain of a glycoprotein (GP) of Marburg virus, and
iii) a third nucleic acid encoding a Marburg virus VP40 matrix protein comprising the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence at least 95% identical thereto;
wherein the first nucleic acid, the second nucleic acid sequence, and the third nucleic acid sequence are each under the control of a promoter compatible with poxvirus expression systems.
14. The recombinant MVA viral vector of claim 13 , wherein the first nucleic acid sequence, the second nucleic acid sequence, and the third nucleic acid sequence are inserted into the MVA in a natural deletion site, a modified natural deletion site, between essential MVA genes, or between non-essential MVA genes.
15. The recombinant MVA viral vector of claim 13 , wherein the first nucleic acid sequence is inserted between MVA genes I8R and G1L or between MVA genes A50R and B1R.
16. The recombinant MVA viral vector of claim 13 , wherein the second nucleic acid sequence is inserted between MVA genes I8R and G1L or between MVA genes A50R and B1R.
17. The recombinant MVA viral vector of claim 13 , wherein the third nucleic acid sequence is inserted between MVA gene site I8R and G1L or between MVA genes site A50R and B1R.
18. The recombinant MVA viral vector of claim 13 , wherein each promoter is selected from the group consisting of a Pm2H5 promoter, a Psyn II promoter, and a mH5 promoter.
19. A pharmaceutical composition comprising at least one recombinant MVA viral vector of claim 1 and a pharmaceutically acceptable adjuvant.
20. The pharmaceutical composition of claim 19 , wherein the adjuvant is an immune response enhancer.
21. A method of inducing an immune response in a subject in need thereof comprising administering an effective amount to the subject the pharmaceutical composition of claim 19 .
22. The method of claim 21 , wherein the subject is a human.