Methods for constructing antibiotic resistance free vaccines
AssigneeThe Trustees of the University of Pennsylvania
InventorsYvonne Paterson, Thorsten Verch
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Abstract
The present invention provides Listeria vaccine strains that express a heterologous antigen and a metabolic enzyme, and methods of generating same.
Claims18
A method of inducing an immune response against a Human Papilloma Virus (HPV) antigen-expressing tumor or a HPV antigen-expressing cancer in a subject, said method comprising the step of administering to the subject a recombinant Listeria strain auxotrophic for D-alanine synthesis, comprising a mutation in a D-alanine racemase gene and in a D-amino acid transferase gene in the chromosome of said recombinant Listeria, wherein said Listeria further comprises a plasmid, wherein the plasmid comprises:
- a first nucleic acid sequence encoding a polypeptide, wherein said polypeptide comprises a heterologous antigen, and
- a second nucleic acid sequence encoding a D-alanine racemase enzyme, wherein said plasmid does not confer antibiotic resistance upon said recombinant Listeria strain,
- whereby said D-alanine racemase gene complements said mutation in said chromosome of said recombinant Listeria, wherein said plasmid comprises a prfA gene, wherein said plasmid is stably maintained in said recombinant Listeria strain in the absence of an antibiotic selection, and wherein said first nucleic acid sequence is operably linked to a prokaryotic promoter/regulatory sequence, wherein said administering induces an immune response against said HPV antigen-expressing tumor or said HPV antigen-expressing cancer in said subject.
A method of regressing a Human Papilloma Virus (HPV) antigen-expressing tumor or an HPV antigen-expressing cancer in a subject, said method comprising the step of administering to the subject a recombinant Listeria strain auxotrophic for D-alanine synthesis, comprising a mutation in a D-alanine racemase gene and in a D-amino acid transferase gene in the chromosome of said recombinant Listeria, wherein said Listeria further comprises a plasmid, wherein the plasmid comprises:
- a first nucleic acid sequence encoding a polypeptide, wherein said polypeptide comprises a heterologous antigen, and
- a second nucleic acid sequence encoding a D-alanine racemase enzyme, wherein said plasmid does not confer antibiotic resistance upon said recombinant Listeria strain,
- whereby said D-alanine racemase gene complements said mutation in said chromosome of said recombinant Listeria, wherein said plasmid comprises prfA gene, wherein said plasmid is stably maintained in said recombinant Listeria strain in the absence of an antibiotic selection, and wherein said first nucleic acid sequence is operably linked to a prokaryotic promoter/regulatory sequence, wherein said administering regresses said HPV antigen expressing tumor or said HPV antigen-expressing cancer in said subject.
Dependencies are inferred from the claim text.