Large and small t antigens of merkel cell polyomavirus, nucleic acid constructs and vaccines made therefrom, and methods of using same

Abstract

Nucleic acid molecules and compositions comprising one or more nucleotide sequences that encode a consensus Merkel Cell Polyomavirus (MCV) T antigen. Immunomodulatory methods and methods of inducing an immune response against MCV are disclosed. Method of treating infection by MCV and methods of treating or preventing Merkel Cell Carcinoma associated with MCV are disclosed. Modified consensus MCV T antigens are disclosed.

1 - 30 . (canceled) 
     
     
         31 . A nucleic acid molecule encoding at least one modified Merkel Cell Polyomavirus (MCV) T antigen, wherein the T antigen comprises at least one mutation that disrupts at least one oncogenic feature of a native MCV T antigen. 
     
     
         32 . The nucleic acid molecule of  claim 31 , wherein the at least one oncogenic feature is selected from the group consisting of CR1 binding, DnaJ binding, phophatase pp2A-binding binding, Rb binding, ATPase activity, helicase activity, chaperone protein binding, hVam6p binding, Fbxw7 binding, origin binding, and transformation. 
     
     
         33 . The nucleic acid molecule of  claim 31 , wherein at least one mutation is a mutation at an amino acid selected from the group consisting of D44, W209, E216, L142, L91, K92, D93, Y94 and M95. 
     
     
         34 . The nucleic acid molecule of  claim 31 , wherein at least one mutation is selected from the group consisting of a D44N mutation, a W209A, an E216K mutation, an L142A mutation, an L91A mutation, a K92A mutation, a D93A mutation, a Y94A mutation and a M95A mutation. 
     
     
         35 . The nucleic acid molecule of  claim 31 , wherein the MCV T antigen is selected from the group consisting of a large T antigen (LTAg), a small t antigen (STAg), and a combination thereof. 
     
     
         36 . The nucleic acid molecule of  claim 31 , encoding at least one peptide comprising an amino acid sequence selected from the group consisting of 
 a) an amino acid sequence having at least about 90% identity over an entire length of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6,   b) an immunogenic fragment comprising at least about 90% identity over at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6,   c) the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6, and   d) an immunogenic fragment comprising at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6.   
     
     
         37 . The nucleic acid molecule of  claim 31 , wherein the nucleic acid molecule is selected from the group consisting of a DNA molecule and an RNA molecule. 
     
     
         38 . The nucleic acid molecule of  claim 31 , wherein the nucleic acid molecule comprises at least one nucleotide sequence selected from the group consisting of 
 a) a nucleotide sequence having at least about 80% identity over an entire length of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:3 and SEQ ID NO:5,   b) an immunogenic fragment of a nucleotide sequence having at least about 80% identity over at least 60% of the nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:3 and SEQ ID NO:5, and   c) an immunogenic fragment comprising at least 60% of the nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:3 and SEQ ID NO:5.   
     
     
         39 . The nucleic acid molecule of  claim 31 , wherein the encoded peptide is operably linked to at least one regulatory sequence selected from the group consisting of a start codon, an IgE leader sequence and a stop codon. 
     
     
         40 . The nucleic acid molecule of  claim 39 , wherein the nucleic acid molecule encodes at least one peptide comprising an amino acid sequence selected from the group consisting of
 a) an amino acid sequence having at least about 90% identity over an entire length of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6,   b) an immunogenic fragment comprising at least about 90% identity over at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6,   c) the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6, and   d) an immunogenic fragment comprising at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6,
 operably linked to an amino acid sequence as set forth in SEQ ID NO:7. 
   
     
     
         41 . The nucleic acid molecule of  claim 39 , wherein the nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of 
 a) a nucleotide sequence having at least about 80% identity over an entire length of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:4 and SEQ ID NO:5,   b) an immunogenic fragment of a nucleotide sequence having at least about 80% identity over at least 60% of the nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:3 and SEQ ID NO:5,and   c) an immunogenic fragment comprising at least 60% of the nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:3 and SEQ ID NO:5,
 operably linked to an nucleotide sequence encoding SEQ ID NO:7. 
   
     
     
         42 . The nucleic acid molecule of  claim 31 , wherein the nucleic acid molecule comprises an expression vector. 
     
     
         43 . The nucleic acid molecule of  claim 31 , wherein the nucleic acid molecule comprises a viral particle. 
     
     
         44 . An immunogenic composition comprising at least one nucleic acid molecule of  claim 31 . 
     
     
         45 . The immunogenic composition of  claim 44 , further comprising at least one selected from the group consisting of a pharmaceutically acceptable excipient and an adjuvant. 
     
     
         46 . A method of treating or preventing a MCV associated pathology in subject in need thereof, the method comprising administering to the subject a peptide comprising an amino acid sequence selected from the group consisting of 
 a) an amino acid sequence having at least about 90% identity over an entire length of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6,   b) an immunogenic fragment comprising at least about 90% identity over at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6,   c) the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6, and   d) an immunogenic fragment comprising at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6.   
     
     
         47 . A method of treating or preventing a MCV associated pathology in subject in need thereof, the method comprising administering to the subject an immunogenic composition comprising a peptide comprising an amino acid sequence selected from the group consisting of 
 a) an amino acid sequence having at least about 90% identity over an entire length of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6,   b) an immunogenic fragment comprising at least about 90% identity over at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID N0:6,   c) the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO: 4 and SEQ ID NO:6, and   d) an immunogenic fragment comprising at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6.   
     
     
         48 . A method of inducing an immune response against a MCV T antigen in a subject in need thereof, the method comprising administering an immunogenic composition of  claim 44  to the subject. 
     
     
         49 . A method of treating or preventing a MCV associated pathology in subject in need thereof, the method comprising administering an immunogenic composition of  claim 31  to the subject. 
     
     
         50 . The method of  claim 49 , wherein the MCV associated pathology is at least one of MCV infection and Merkel Cell Carcinoma.