Dual delivery system for heterologous antigens
Provided herein are recombinant Listeria strains expressing a tumor-specific antigenic polypeptide and, optionally, an angiogenic polypeptide wherein a nucleic acid molecule encoding at least one of the polypeptides is operably integrated into the Listeria genome in an open reading frame with a nucleic acid sequence encoding a PEST-containing polypeptide, methods of preparing same, and methods of inducing an immune response, and treating, inhibiting, or suppressing cancer or tumors comprising administering same.
1. A recombinant Listeria strain comprising a first and second nucleic acid molecule, each said nucleic acid molecule encoding a heterologous antigenic polypeptide or immunogenic fragment thereof, wherein said first nucleic acid molecule is operably integrated into the Listeria genome in an open reading frame with a first endogenous PEST-containing gene, and wherein said recombinant Listeria strain comprises an episomal expression vector comprising said second nucleic acid molecule encoding a heterologous antigen operably fused to a second PEST-containing gene, wherein said Listeria strain is a recombinant dal/dat mutant strain, wherein said Listeria strain comprises a mutation or deletion of said Listeria endogenous actA gene.
2. The recombinant Listeria of claim 1 , wherein said episomal expression vector comprises an open reading frame encoding a metabolic enzyme for complementing said dal/dat mutant.
3. The recombinant Listeria strain of claim 1 , wherein said first PEST-containing gene is LLO.
4. The recombinant Listeria strain of claim 1 , wherein said first PEST-containing gene is ActA.
5. The recombinant Listeria strain of claim 1 , wherein said first or second nucleic acid molecule encodes a polypeptide expressed by a tumor cell.
6. The recombinant Listeria strain of claim 1 , wherein said first or second nucleic acid molecule encodes an angiogenic polypeptide.
7. The recombinant Listeria strain of claim 1 , wherein said first or second nucleic acid molecule encodes a prostate specific antigen (PSA).
8. The recombinant Listeria strain of claim 1 , wherein said first or second nucleic acid molecule encodes a High Molecular Weight-Melanoma Associated Antigen (HMW-MAA).
9. The recombinant Listeria strain of claim 1 , wherein said first or second nucleic acid molecule encodes a chimeric Her2 antigen (cHer2).
10. The recombinant Listeria strain of claim 1 , wherein said first nucleic acid molecule is a vector designed for site-specific homologous recombination into the Listeria genome.
11. The recombinant Listeria strain of claim 1 , wherein said recombinant Listeria strain is an auxotrophic Listeria strain.
12. The recombinant Listeria strain of claim 2 , wherein said metabolic enzyme is an alanine racemase enzyme.
13. The recombinant Listeria strain of claim 2 , wherein said metabolic enzyme is a D-amino acid transferase enzyme.
14. The recombinant Listeria strain of claim 1 , wherein said recombinant Listeria strain has been passaged through an animal host.
15. The recombinant Listeria strain of claim 1 , wherein said recombinant Listeria strain is a recombinant Listeria monocytogenes strain.
16. An immunogenic composition comprising the recombinant Listeria strain of claim 1 and an adjuvant, cytokine, chemokine, or combination thereof.
17. The recombinant Listeria strain of claim 1 , wherein said first or second nucleic acid molecule encodes a carbonic anhydrase 9 (CA9) antigen or fragment thereof.
18. The recombinant Listeria strain of claim 1 , wherein said second PEST-containing gene comprises a truncated LLO (tLLO), a truncated ActA, or a PEST sequence peptide.