Antigen delivery platforms
This disclosure provides platforms for delivery of herpes virus proteins to cells, particularly proteins that form complexes in vivo. In some embodiments these proteins and the complexes they form elicit potent neutralizing antibodies. Thus, presentation of herpes virus proteins using the disclosed platforms permits the generation of broad and potent immune responses useful for vaccine development.
1. A self-replicating RNA molecule comprising a polynucleotide which comprises:
a) a first nucleotide sequence encoding a first protein or fragment thereof from cytomegalovirus (CMV), wherein the first nucleotide sequence is operably linked to a subgenomic promoter and followed by b);
b) a second nucleotide sequence encoding a second protein or fragment thereof from said CMV, wherein the second nucleotide sequence is operably linked to a subgenomic promoter and followed by c);
c) a third nucleotide sequence encoding a third protein or fragment thereof from said CMV, wherein the third nucleotide sequence is operably linked to a subgenomic promoter and followed by d);
d) a fourth nucleotide sequence encoding a fourth protein or fragment thereof from said CMV, wherein the fourth nucleotide sequence is operably linked to an IRES or a viral 2A site and followed by e); and
e) a fifth nucleotide sequence encoding a fifth protein or fragment thereof from said CMV, wherein the fifth nucleotide sequence is operably linked to an IRES or a viral 2A site,
wherein the first protein is gH, the second protein is gL, the third protein is UL128, the fourth protein is UL130, and the fifth protein is UL131;
and wherein introducing the self-replicating RNA molecule into a suitable cell results in the expression of the first, second, third, fourth and fifth CMV proteins or fragments thereof in an amount sufficient for the formation of a gH/gL/UL128/UL130/UL131 pentameric complex.
2. The self-replicating RNA molecule of claim 1 , wherein the first protein consists of SEQ ID NO: 32 or a fragment thereof.
3. The self-replicating RNA molecule of claim 1 , wherein the second protein consists of SEQ ID NO: 36 or a fragment thereof.
4. The self-replicating RNA molecule of claim 1 , wherein the third protein consists of SEQ ID NO: 44 or a fragment thereof.
5. The self-replicating RNA molecule of claim 1 , wherein the fourth protein consists of SEQ ID NO: 46 or a fragment thereof.
6. The self-replicating RNA molecule of claim 1 , wherein the fifth protein consists of SEQ ID NO: 48 or a fragment thereof.
7. The self-replicating RNA molecule of claim 1 , wherein the first protein consists of SEQ ID NO: 32 or a fragment thereof; the second protein consists of SEQ ID NO: 36 or a fragment thereof; the third protein consists of SEQ ID NO: 44 or a fragment thereof; the fourth protein consists of SEQ ID NO: 46 or a fragment thereof; and the fifth protein consists of SEQ ID NO: 48 or a fragment thereof.
8. The self-replicating RNA molecule of claim 7 , wherein the self-replicating RNA molecule is encoded by a DNA sequence selected from the group consisting of SEQ ID NO: 56 (vector A526) and SEQ ID NO: 57 (vector A527).
9. The self-replicating RNA molecule of claim 1 , wherein the self-replicating RNA molecule is an alphavirus replicon.
10. The self-replicating RNA molecule of claim 1 , wherein the subgenomic promoter comprises SEQ ID NO:51.
11. The self-replicating RNA molecule of claim 1 , wherein the IRES comprises SEQ ID NO:49 or SEQ ID NO:50.
12. The self-replicating RNA molecule of claim 1 , wherein the viral 2A site comprises SEQ ID NO:2.
13. The self-replicating RNA molecule of claim 12 , wherein the viral 2A site comprises SEQ ID NO:3.
14. A composition comprising the self-replicating RNA of claim 1 and an RNA delivery system.
15. The composition of claim 14 , wherein the RNA delivery system is a liposome, a polymeric nanoparticle, a lipid nanoparticle (LNP), an oil-in-water cationic nanoemulsion or combinations thereof.
16. A method of inducing an immune response in an individual, comprising administering to the individual a composition of claim 14 .
17. A recombinant DNA molecule that encodes the self-replicating RNA molecule of claim 1 .
18. The recombinant DNA molecule of claim 17 , wherein the recombinant DNA molecule is a plasmid.
19. The recombinant DNA molecule of claim 18 , wherein the recombinant DNA molecule comprises a DNA sequence selected from the group consisting of SEQ ID NO: 56 (vector A526) and SEQ ID NO: 57 (vector A527).