Multi-epitopic vaccine
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.
1. An isolated polypeptide comprising:
(i) at least one CD4 + epitope(s);
(ii) at least one CD8 + epitope(s), wherein said 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
(iii) a protein transduction domain consisting of SEQ ID NO: 8 or a protein transduction domain consisting of amino acids 1 to 43 of SEQ ID NO: 8.
2. The isolated polypeptide according to claim 1 , wherein the polypeptide comprises a protein transduction domain consisting of SEQ ID NO: 8, at least one CD4 + epitope(s), and at least one CD8 + epitope(s), wherein said 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.
3. The isolated polypeptide according to claim 1 , wherein said polypeptide contains at least two CD4 + epitopes and at least two CD8 + epitopes and:
(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.
4. Isolated antigen-presenting cells loaded with the polypeptide according to claim 1 .
5. The isolated antigen presenting cells according to claim 4 , which are selected from the group consisting of dendritic cells, macrophages and B-cells.
6. 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.
7. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.
8. A method of preparing a pharmaceutical composition comprising the step of mixing the polypeptide of claim 1 and a pharmaceutically acceptable carrier.
9. The isolated polypeptide according to claim 1 , wherein said CD4 + epitope consists of about 8-25 amino acids and said CD8 + epitope consists of about 8-15 amino acids.
10. The isolated polypeptide according to claim 1 , wherein said CD4 + epitope consists of about 12 to about 25 amino acids and said CD8 + epitope consists of about 8 to about 11 amino acids.
11. The isolated polypeptide according to claim 1 , wherein the polypeptide comprises a protein transduction domain consisting of amino acids 1 to 43 of SEQ ID NO: 8, at least one CD4 + epitope(s), and at least one CD8 + epitope(s), wherein said 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.
12. The isolated polypeptide according to claim 1 , wherein the polypeptide consists of (i) at least one CD4 + epitope(s), (ii) at least one CD8 + epitope(s), wherein said 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 (iii) a protein transduction domain consisting of the amino acid sequence SEQ ID NO: 8, or amino acids 1 to 43 of SEQ ID NO: 8.
13. The isolated polypeptide according to claim 12 , wherein the polypeptide consists of (i) at least one CD4 + epitope(s), (ii) at least one CD8 + epitope(s), wherein said 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 (iii) a protein transduction domain consisting of amino acids 1 to 43 of SEQ ID NO: 8.
14. The isolated polypeptide according to claim 12 , wherein the polypeptide consists of (i) at least one CD4 + epitope(s), (ii) at least one CD8 + epitope(s), wherein said 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 a protein transduction domain consisting of the amino acid sequence SEQ ID NO: 8.