Compositions and methods for treatment of cervical dysplasia

Abstract

The present invention provides methods of treating, protecting against, and inducing an immune response against cervical dysplasia and cancer, comprising the step of administering to a subject a recombinant Listeria strain, comprising a fusion peptide that comprises an LLO fragment and an E7 and/or E6 antigen. The present invention also provides methods for inducing an anti-E7 CTL response in a human subject and treating HPV-mediated diseases, disorders, and symptoms, comprising administration of the recombinant Listeria strain.

What is claimed: 
     
         1 . A polynucleotide encoding a fusion protein comprising:
 a. a PEST-like sequence from a prokaryotic organism; and   b. a tumor antigen.   
     
     
         2 . The polynucleotide of  claim 1 , wherein the prokaryotic organism is a  Listeria  bacterium. 
     
     
         3 . The polynucleotide of  claim 2 , wherein the  Listeria  bacteria is  Listeria monocytogenes.    
     
     
         4 . The polynucleotide of  claim 1 , wherein the PEST-like sequence is selected from the sequences set forth in SEQ ID NOs: 2-5. 
     
     
         5 . The polynucleotide of  claim 1 , wherein the polynucleotide is operably linked to a promoter. 
     
     
         6 . The polynucleotide of  claim 5 , wherein the promoter is selected from the group consisting of Listerial prfA, Listerial hly promoter, Listerial actA promoter, and Listerial p60 promoter. 
     
     
         7 . The polynucleotide of  claim 1 , wherein the tumor antigen is a self-antigen. 
     
     
         8 . A method of enhancing immunogenicity of a tumor antigen in a  Listeria  bacteria, the method comprising introducing a polynucleotide into the  Listeria  bacteria, wherein the polynucleotide encodes a fusion protein comprising the tumor antigen and a PEST-like sequence from the  Listeria  bacteria. 
     
     
         9 . The method of  claim 8 , wherein the  Listeria  bacteria is  Listeria monocytogenes.    
     
     
         10 . The method of  claim 8 , wherein the PEST-like sequence is selected from the sequences set forth in SEQ ID NOs: 2-5. 
     
     
         11 . The method of  claim 8 , wherein the polynucleotide is operably linked to a promoter. 
     
     
         12 . The method of  claim 11 , wherein the promoter is selected from the group consisting of Listerial prfA promoter, Listerial hly promoter, Listerial actA promoter, and Listerial p60 promoter. 
     
     
         13 . The method of  claim 8 , wherein the tumor antigen is a self-antigen. 
     
     
         14 . The method of  claim 8 , wherein the  Listeria  bacteria is  Listeria monocytogenes.    
     
     
         15 . A polynucleotide encoding a fusion protein comprising:
 c. a fragment of ActA protein from a  Listeria  bacterium, wherein the fragment of ActA protein comprises one or more PEST-like sequences; and   d. a tumor antigen.   
     
     
         16 . The polynucleotide of  claim 15 , wherein the polynucleotide is operably linked to a promoter. 
     
     
         17 . The polynucleotide of  claim 16 , wherein the promoter is selected from the group consisting of Listerial prfA promoter, Listerial hly promoter, Listerial actA promoter, and Listerial p60 promoter. 
     
     
         18 . The polynucleotide of  claim 15 , wherein the fragment of ActA protein has an amino acid sequence at least 80% homologous with the amino acid sequence set forth in SEQ ID NO: 23. 
     
     
         19 . The polynucleotide of  claim 15 , wherein the fragment of ActA protein has an amino acid sequence as set forth in SEQ ID NO: 23. 
     
     
         20 . The polynucleotide of 15, wherein the PEST-like sequences are selected from the sequences set forth in SEQ ID NOs: 2-5. 
     
     
         21 . The polynucleotide of  claim 15 , wherein the tumor antigen is a self-antigen. 
     
     
         22 . A method of enhancing immunogenicity of a tumor antigen in a first  Listeria  bacterium, the method comprising introducing a polynucleotide into the first  Listeria  bacterium, wherein the polynucleotide encodes a fusion protein comprising the tumor antigen and a fragment of ActA protein from a second  Listeria  bacterium, wherein the fragment of ActA protein comprises one or more PEST-like sequences. 
     
     
         23 . The method of  claim 22 , wherein the polynucleotide is operably linked to a promoter. 
     
     
         24 . The method of  claim 23 , wherein the promoter is selected from the group consisting of Listerial prfA promoter, Listerial hly promoter, Listerial actA promoter, and Listerial p60 promoter. 
     
     
         25 . The method of  claim 22 , the fragment of ActA protein has an amino acid sequence at least 80% homologous with the amino acid sequence set forth in SEQ ID NO: 23. 
     
     
         26 . The method of  claim 22 , wherein the fragment of ActA protein has an amino acid sequence as set forth in SEQ ID NO: 23. 
     
     
         27 . The method of  claim 22 , wherein the tumor antigen is a self-antigen. 
     
     
         28 . The method of  claim 22 , wherein the first  Listeria  bacterium and the second  Listeria  bacterium belong to the same subtype of  Listeria monocytogenes.    
     
     
         29 . The method of  claim 23 , wherein the first  Listeria  bacterium and the second  Listeria  bacterium belong to different subtypes of  Listeria monocytogenes.    
     
     
         30 . The method of  claim 22 , wherein the PEST-like sequences are selected from the sequences set forth in SEQ ID NOs: 2-5.