Altering the immundominance hierarchy using a DNA vaccine expressing conserved regions
The invention provides methods and compositions for eliciting broad immune responses. The methods employ nucleic acid vaccines that encodes highly conserved elements from a virus.
1. A nucleic acid comprising a nucleotide sequence encoding a fusion polypeptide comprising the conserved element amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7, wherein the conserved elements are separated by alanine-containing peptide linker amino acid sequences.
2. The nucleic acid of claim 1 , wherein the fusion polypeptide comprises the amino acid sequence SEQ ID NO:15.
3. The nucleic acid of claim 2 , comprising the region of the nucleic acid sequence of SEQ ID NO:17 that encodes p24CE1.
4. The nucleic acid of claim 2 , wherein the fusion polypeptide comprises a GM-CSF signal peptide.
5. The nucleic acid of claim 4 , wherein the fusion polypeptide comprises the amino acid sequence of SEQ ID NO:18.
6. The nucleic acid of claim 2 , wherein the fusion polypeptide comprises the amino acid sequence of SEQ ID NO:23 or the amino acid sequence of SEQ ID NO:27.
7. The nucleic acid of claim 1 , wherein the nucleic acid is an RNA molecule.
8. The nucleic acid of claim 1 , wherein the nucleic acid is a DNA molecule.
9. An expression vector comprising the nucleic acid sequence of claim 1 .
10. The expression vector of claim 9 , further comprising a nucleic acid encoding a second fusion polypeptide comprising the conserved element amino acid sequences of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14 separated by alanine-containing peptide linker amino acid sequences.
11. The expression vector of claim 10 , wherein the second fusion polypeptide comprises the amino acid sequence SEQ ID NO:16.
12. The expression vector of claim 11 , wherein the nucleic acid encoding the second fusion polypeptide comprises the region of the nucleic acid sequence of SEQ ID NO:17 that encodes p24CE2.
13. The expression vector of claim 11 , wherein the second fusion polypeptide comprises a GM-CSF signal peptide.
14. The expression vector of claim 13 , wherein the second fusion polypeptide comprises the amino acid sequence of SEQ ID NO:19.
15. The expression vector of claim 11 , wherein the second fusion polypeptide comprises the amino acid sequence of SEQ ID NO:21 or comprises the amino acid sequence of SEQ ID NO:29.
16. A nucleic acid construct comprising the nucleic acid of claim 1 , and further comprising a nucleic acid encoding a second fusion polypeptide comprising the conserved element amino acid sequences of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14, wherein the conserved elements are separated by alanine-containing peptide linker amino acid sequences.
17. A nucleic acid construct comprising the nucleic acid of claim 2 , further comprising a nucleic acid encoding a second fusion polypeptide comprising the amino acid sequence of SEQ ID NO:16.
18. The nucleic acid construct of claim 17 , comprising the nucleic acid sequence of SEQ ID NO:17.
19. The nucleic acid construction of claim 17 , wherein the construct is an RNA molecule.