Modified vp1-capsid protein of parvovirus b19

Abstract

The inventions relates to a modified VP1-capsid protein of parvovirus B19 having reduced phospholipase A2-like enzyme activity as compared to the wild type VP1-capsid protein of parvovirus B19 having the amino acid sequence of SEQ ID NO: 1.

1 .- 41 . (canceled) 
     
     
         42 . A modified VP1-capsid protein of parvovirus B19, wherein the wild type sequence of the VP-1 capsid protein has been modified to contain amino acid substitutions at positions corresponding to histidine 153 of SEQ ID NO:1, tyrosine 157 of SEQ ID NO:1, lysine 162 of SEQ ID NO:1, and tyrosine 168 of SEQ ID NO:1, wherein said substitutions are histidine 153 to alanine, tyrosine 157 to phenylalanine, lysine 162 to leucine, and tyrosine 168 to phenylalanine, and wherein said modified VP1-capsid protein has a reduced phospholipase A2 enzyme activity as compared to the wild type VP1-capsid protein. 
     
     
         43 . A method of treating parvovirus B19 infection comprising administering to a human subject a pharmaceutical composition comprising:
 (a) an adjuvant, and   (b) a modified VP1-capsid protein of parvovirus B19 wherein the wild type sequence of the VP-1 capsid protein has been modified to contain amino acid substitutions at positions corresponding to histidine 153 of SEQ ID NO:1, tyrosine 157 of SEQ ID NO:1, lysine 162 of SEQ ID NO:1, and tyrosine 168 of SEQ ID NO:1, wherein said substitutions are histidine 153 to alanine, tyrosine 157 to phenylalanine, lysine 162 to leucine, and tyrosine 168 to phenylalanine, and wherein said modified VP1-capsid protein has a reduced phospholipase A2 enzyme activity as compared to the wild type VP1-capsid protein.   
     
     
         44 . The method of  claim 43 , wherein said adjuvant is an immunostimulatory substance selected from the group consisting of an immunostimulatory deoxynucleotide (ODN), a peptide containing at least two LysLeuLys motifs, a neuroactive compound, alum, Freund's complete adjuvant and Freund's incomplete adjuvant. 
     
     
         45 . The method of  claim 43 , further comprising a polycationic peptide. 
     
     
         46 . The method of  claim 43 , further comprising a VP2-capsid protein. 
     
     
         47 . The method of  claim 46 , wherein the modified VP1-capsid protein is fused to the VP2-capsid protein. 
     
     
         48 . A method of inducing an immune response against parvovirus B19 infection in a human, comprising administering to a human subject a pharmaceutical composition comprising:
 (a) an adjuvant, and   (b) a modified VP1-capsid protein of parvovirus B19 wherein the wild type sequence of the VP-1 capsid protein has been modified to contain amino acid substitutions at positions corresponding to histidine 153 of SEQ ID NO:1, tyrosine 157 of SEQ ID NO:1, lysine 162 of SEQ ID NO:1, and tyrosine 168 of SEQ ID NO:1, wherein said substitutions are histidine 153 to alanine, tyrosine 157 to phenylalanine, lysine 162 to leucine, and tyrosine 168 to phenylalanine, and wherein said modified VP1-capsid protein has a reduced phospholipase A2 enzyme activity as compared to the wild type VP1-capsid protein.   
     
     
         49 . The method of  claim 48 , wherein said immune response provides protection against human parvovirus B19 infection. 
     
     
         50 . The method of  claim 48 , wherein said adjuvant is an immunostimulatory substance selected from the group consisting of an immunostimulatory deoxynucleotide (ODN), a peptide containing at least two LysLeuLys motifs, a neuroactive compound, alum, Freund's complete adjuvant and Freund's incomplete adjuvant. 
     
     
         51 . The method of  claim 48 , further comprising a polycationic peptide. 
     
     
         52 . The method of  claim 48 , further comprising a VP2-capsid protein. 
     
     
         53 . The method of  claim 52 , wherein the modified VP1-capsid protein is fused to the VP2-capsid protein.