Antigen delivery platforms

Abstract

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 a herpes virus or fragment thereof; and   b. a second nucleotide sequence encoding a second protein or fragment thereof from said herpes virus or fragment thereof;   wherein the first nucleotide sequence and second nucleotide sequence are operably linked to one or more control elements so that when the self-replicating RNA molecule is introduced into a suitable cell, the first and second herpes virus proteins or fragments thereof are produced in an amount sufficient for the formation of a complex in the cell that contains the first and second proteins or fragments.   
     
     
         2 . The self-replicating RNA molecule of  claim 1  with the proviso that the first protein and the second protein are not the same protein or fragments of the same protein, the first protein is not a fragment of the second protein, and the second protein is not a fragment of the first protein. 
     
     
         3 . The self-replicating RNA molecule of  claim 2 , wherein the first nucleotide sequence is operably linked to a first control element and the second nucleotide sequence is operably linked to a second control element. 
     
     
         4 . The self-replicating RNA molecule of  claim 1 , further comprising a third nucleotide sequence encoding a third protein or fragment thereof from said herpes virus, wherein the third nucleotide sequences is operably linked to a control element. 
     
     
         5 . The self-replicating RNA molecule of  claim 4 , wherein the third nucleotide sequence is operably linked to a third control element. 
     
     
         6 . The self-replicating RNA molecule of  claim 4 , further comprising a fourth nucleotide sequence encoding a fourth protein or fragment thereof from said herpes virus, wherein the fourth nucleotide sequences is operably linked to a control element. 
     
     
         7 . The self-replicating RNA molecule of  claim 6 , wherein the fourth nucleotide sequence is operably linked to a fourth control element. 
     
     
         8 . The self-replicating RNA molecule of  claim 6 , further comprising a fifth nucleotide sequence encoding a fifth protein or fragment thereof from said herpes virus, wherein the fifth nucleotide sequences is operably linked to a control element. 
     
     
         9 . The self-replicating RNA molecule of  claim 8 , wherein the fifth nucleotide sequence is operably linked to a fifth control element. 
     
     
         10 . The self-replicating RNA molecule of  claim 1  wherein the control elements are independently selected from the group consisting of a subgenomic promoter, an IRES, and a viral 2A site. 
     
     
         11 . The self-replicating RNA molecule of  claim 1 , wherein the herpes virus is cytomegalovirus (CMV). 
     
     
         12 . The self-replicating RNA molecule of  claim 8  wherein the first protein or fragment, the second protein or fragment, the third protein or fragment, the fourth protein or fragment, and the fifth protein or fragment are independently selected from the group consisting of gB, gH, gL; gO; gM, gN; UL128, UL130, UL131, and a fragment of any one of the foregoing. 
     
     
         13 . The self-replicating RNA molecule of  claim 11 , wherein the first protein or fragment is gH or a fragment thereof, and the second protein or fragment is gL or a fragment thereof. 
     
     
         14 . The self-replicating RNA molecule of  claim 11 , wherein the first protein or fragment is gH or a fragment thereof, and the second protein or fragment is gL or a fragment thereof, and the third protein or fragment is gO or a fragment thereof. 
     
     
         15 . The self-replicating RNA molecule of  claim 11 , wherein the first protein or fragment is gH or a fragment thereof, and the second protein or fragment is gL or a fragment thereof, the third protein or fragment is UL128 or a fragment thereof, the fourth protein or fragment is UL130 or a fragment thereof, and the fifth protein or fragment is UL131 or a fragment thereof. 
     
     
         16 . The self-replicating RNA molecule of  claim 1 , wherein the herpes virus is varicella zoster virus (VZV). 
     
     
         17 . The self-replicating RNA molecule of  claim 16 , wherein the first protein or fragment, the second protein or fragment, the third protein or fragment, the fourth protein or fragment, and the fifth protein or fragment are independently selected from the group consisting of gB, gE, gH, gI, gL, and a fragment of any one of the foregoing. 
     
     
         18 . The self-replicating RNA molecule of  claim 16 , wherein the first protein or fragment is gH or a fragment thereof, and the second protein or fragment is gL or a fragment thereof. 
     
     
         19 . The self-replicating RNA molecule of  claim 8 , wherein the self-replicating RNA molecule is an alphavirus replicon. 
     
     
         20 . (canceled) 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . A composition comprising the self-replicating RNA of  claim 1  and a pharmaceutically acceptable vehicle. 
     
     
         25 . The composition of  claim 24 , further comprising a second self-replicating RNA molecule that encodes a protein or fragment thereof from the herpes virus. 
     
     
         26 . The composition of  claim 24 , further comprising an RNA delivery system. 
     
     
         27 . The composition of  claim 26 , wherein the RNA delivery system is a liposome, a polymeric nanoparticle, an oil-in-water cationic nanoemulsion or combinations thereof. 
     
     
         28 . (canceled) 
     
     
         29 . A method of forming a protein complex, comprising delivering an alphavirus replicon particle (VRP) comprising a self-replicating RNA molecule of  claim 8  to a cell, and maintaining the cell under conditions suitable for expression of the alphavirus replicon, wherein a protein complex is formed. 
     
     
         30 . The method of  claim 29  wherein the cell is in vivo. 
     
     
         31 . A method of forming a protein complex, comprising delivering the self-replicating RNA of  claim 1  to a cell, and maintaining the cell under conditions suitable for expression of said self-replicating RNA, wherein a protein complex is formed. 
     
     
         32 . (canceled) 
     
     
         33 . A method of inhibiting herpes virus entry into a cell comprising contacting the cell with the self-replicating RNA of  claim 1 . 
     
     
         34 . A method of inhibiting herpes virus entry into a cell comprising contacting the cell with an alphavirus replicon particle (VRP) comprising a self-replicating RNA molecule of  claim 8 . 
     
     
         35 . The method of  claim 33  wherein the cell is selected from the group consisting of an epithelial cell, an endothelial cell, and a fibroblast. 
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . (canceled) 
     
     
         39 . A method of inducing an immune response in an individual, comprising administering to the individual a self-replicating RNA molecule of  claim 1 . 
     
     
         40 . The method of  claim 39 , wherein the immune response comprises the production of neutralizing antibodies. 
     
     
         41 . The method of  claim 40  wherein the neutralizing antibodies are complement-independent. 
     
     
         42 . A recombinant DNA molecule that encodes the self-replicating RNA molecule of  claim 1 . 
     
     
         43 . The recombinant DNA molecule of  claim 42 , wherein the recombinant DNA molecule is a plasmid. 
     
     
         44 . (canceled) 
     
     
         45 . (canceled) 
     
     
         46 . (canceled)