Multi-epitopic vaccine

Abstract

The present invention relates to isolated polypeptides comprising: (i) a protein transduction domain consisting of ZEBRA or a fragment thereof that retains the capacity of internalization, (ii) at least one CD4 + epitope; and (iii) at least one CD8 + epitope. It also relates to antigen presenting cells loaded with said polypeptides, and the use thereof in immunotherapy including prevention and/or treatment of cancers or infectious diseases.

We claim: 
     
         1 . An isolated polypeptide comprising:
 (i) a protein transduction domain consisting of ZEBRA consisting of amino acids residues 178-220 of ZEBRA or a fragment thereof or a variant thereof having at least one conservatively substituted amino acid from the native sequence, that retains the capacity of internalization,   (ii) at least one CD4 +  epitope(s); and   (iii) at least one CD8 +  epitope(s).   
     
     
         2 . The isolated polypeptide according to  claim 1 , wherein the protein transduction domain comprises or consists of the amino acid sequence having at least 80%, at least 90% identity with SEQ ID NO: 8. 
     
     
         3 . The isolated polypeptide according to  claim 1 , wherein:
 (i) the CD4 +  and CD8 +  epitopes are selected from the group consisting of epitopes from a tumor-associated antigen, epitopes from a tumor-specific antigen, and epitopes from an antigenic protein from a pathogen; and   (ii) said CD4 +  epitope consists of about 6-100 amino acids and said CD8 +  epitope consists of about 6-100 amino acids.   
     
     
         4 . The isolated polypeptide according to  claim 1 , wherein:
 (i) said at least two CD4 +  epitopes are restricted to at least two MHC class II molecules; and   (ii) said at least two CD8 +  epitopes are restricted to at least two MHC class I molecules of the human population.   
     
     
         5 . An isolated polynucleotide encoding a polypeptide comprising:
 (i) a protein transduction domain consisting of ZEBRA consisting of amino acids residues 178-220 of ZEBRA or a fragment thereof or a variant thereof having at least one conservatively substituted amino acid from the native sequence, that retains the capacity of internalization,   (ii) at least one CD4 +  epitope(s); and   (iii) at least one CD8 +  epitope(s).   
     
     
         6 . The isolated polynucleotide according to  claim 5 , wherein the nucleotide sequence corresponding to the protein transduction domain comprises or consists of any nucleotide sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with SEQ ID NO: 7. 
     
     
         7 . A recombinant vector comprising the polynucleotide of  claim 5 . 
     
     
         8 . A host cell comprising the recombinant vector of  claim 7 . 
     
     
         9 . A method for preparing the polypeptide comprising cultivating the host cell according to  claim 8  in a culture medium and separating said polypeptide from the culture medium or separating said polypeptide from the host cell lysate after host cell lysis. 
     
     
         10 . Antigen-presenting cells loaded with the polypeptide according to  claim 1 . 
     
     
         11 . Antigen presenting cells according to  claim 10 , which are selected among dendritic cells, macrophages and B-cells or dendritic cells. 
     
     
         12 . A method for preparing antigen presenting cells, comprising transducing antigen presenting cells with the polypeptide of  claim 1 , cultivating said cells in a culture medium and separating said cells from the culture medium. 
     
     
         13 . A pharmaceutical composition comprising the polypeptide of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         14 . A method of preparing the pharmaceutical composition comprising the step of mixing the polypeptide of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         15 . A method for eliciting or improving, in a subject, an immunologic response against multiple epitopes that is (i) dependent on CD4 +  helper T cells and CD8 +  cytotoxic T cells and/or (ii) is restricted by multiple MHC class I molecules and multiple MHC class II molecules, wherein said method comprises administering the composition of  claim 13  to said subject. 
     
     
         16 . A method of preventing, treating or stabilizing a disease or disorder in a subject, said method comprising administering, in a subject in need thereof, a therapeutically effective amount of a composition of  claim 13 . 
     
     
         17 . The method of  claim 16 , wherein the disease or disorder is a cancer, an infectious disease, an auto-immunity disorder or a transplant rejection. 
     
     
         18 . The method of  claim 17 , wherein the antigen presenting cells are dendritic cells from the subject to be treated. 
     
     
         19 . The method of  claim 16 , wherein the composition is administered subcutaneously. 
     
     
         20 . The method of  claim 17 , wherein said method is for treating, preventing or stabilizing a cancer and is carried out in combination with chemotherapy, radiotherapy, surgery, targeted therapy (including small molecules, peptides and monoclonal antibodies) and/or anti-angiogenic therapy.