Cellular immunity inducing vaccine

Abstract

A novel vaccine that can induce sufficiently high cell-mediated immunity is disclosed. The vaccine of the present invention contains, as an effective component, a polypeptide comprising a tandem repeat structure in which an MHC class I epitope region derived from an antigen protein and a spacer sequence are linked to each other alternately and repeatedly at least three times, or a recombinant vector which comprises a polynucleotide encoding said polypeptide and is capable of expressing said polypeptide in vivo. The spacer sequence is, for example, a sequence generated as an amino acid sequence inevitably encoded by a single base sequence which is designed such that the MHC class I epitope region derived from the antigen protein, an MHC class II epitope region derived from the antigen protein, and at least one higher-order-structure-stabilizing region are encoded by different reading frames in said single base sequence.

1 . A method of manufacturing a vaccine, said method comprising:
 a designing step of designing a multifunctional base sequence in which an MHC class I epitope region, an MHC class II epitope region and at least one higher-order-structure-stabilizing region are encoded separately by three reading frames, wherein the MHC class I epitope region and the MHC class II epitope region are derived from the same antigen protein or different antigen proteins;   a production step of producing a polypeptide comprising a tandem repeat structure in which the MHC class I epitope region and a spacer sequence are linked to each other alternately and repeatedly at least three times, or a polynucleotide encoding said polypeptide and is capable of expressing said polypeptide in vivo, wherein said spacer sequence is an amino acid sequence that is generated adjacent to the MHC class I epitope region in a reading reading frame encoding the MHC class I epitope region in the multifunctional base sequence, or an amino acid sequence which is the same amino acid sequence as (1) except that several amino acids are substituted; and   a formulation step of mixing the polypeptide or the polynucleotide with a pharmaceutically acceptable additive(s).   
     
     
         2 . The method according to  claim 1 , wherein said higher-order-structure-stabilizing region is at least one selected from the group consisting of an α-helix-forming region, a β-sheet-forming region and hydrophobic-bond-forming region. 
     
     
         3 . The method according to  claim 1 , wherein said MHC class I epitope region and said MHC class II epitope region are derived from the same antigen protein. 
     
     
         4 . The method according to  claim 1 , wherein said polypeptide comprises said MHC class II epitope region in at least one of an N-terminal region and a C-terminal region. 
     
     
         5 . The method according to  claim 1 , wherein the production step comprises polymerizing said multifunctional base sequence to obtain a library of artificial genes, allowing the artificial genes to express proteins, and selecting said polypeptide from the proteins expressed. 
     
     
         6 . The method according to  claim 1 , wherein said antigen protein is a tumor antigen, cancer stem cell antigen, viral antigen, or parasite antigen. 
     
     
         7 . The method according to  claim 1 , wherein said antigen protein is WT1, survivin, survivin-B2, MAGE-A3, MEGE-A4, tyrosinase, gp100, Melan-A, TRP-2, SNRPD1, CDK4, NY-ESO-1, HER2, MUC-1, CD20, or p53. 
     
     
         8 . The method according to  claim 1 , wherein the isoelectric point of said polypeptide is 6.0 to 8.6.