Agents and methods based on the use of the EDA domain of fibronectin
This invention relates to the use of a polypeptide in the production of an immunostimulatory agent, said polypeptide comprising a sequence corresponding to the EDA domain of fibronectin, a fragment of the EDA domain which can bind to TLR4 or a variant of said EDA domain or a fragment which can bind to TLR4 and which has a homology of more than 70% with any form or natural fragment of the EDA domain. The invention also relates to the production methods and applications of said agent.
1. A proteinaceous vector which comprises a polypeptide whose amino acid sequence is selected from:
a) fibronectin EDA domain (EDA), or
b) a fragment of an EDA domain capable of binding TLR4, wherein said EDA domain consists of
residues 2 to 91 of SEQ. ID. NO: 2,
SEQ. ID. NO: 4,
residues 2 to 57 of SEQ. ID. NO: 6, or
SEQ. ID. NO: 8 bound to a molecule of interest selected from the group consisting of: a viral antigen, a fungal antigen, a parasitic antigen, a tumoral antigen, and a tumoral antigenic determinant,
wherein the proteinaceous vector is capable of translocating the molecule of interest into a TLR4 expressing cell.
2. The proteinaceous vector according to claim 1 , wherein said molecule of interest is a viral antigen.
3. The proteinaceous vector according to claim 2 , wherein said molecule of interest is a viral antigen from hepatitis C virus.
4. The proteinaceous vector according to claim 3 , wherein the hepatitis C viral antigen is NS3 protein or an antigenic fragment of said protein.
5. The proteinaceous vector according to claim 4 , wherein said hepatitis C viral antigen consists of the amino acid sequence of SEQ ID NO: 10.
6. The proteinaceous vector according to claim 1 , wherein said molecule of interest is selected from a tumoral antigen and a tumoral antigenic determinant.
7. The proteinaceous vector according to claim 1 , wherein the proteinaceous vector comprises an amino acid sequence selected from the group consisting of amino acid residues 2-91 of SEQ ID NO: 2 and amino acid residues 2-57 of SEQ ID NO: 6.
8. The proteinaceous vector according to claim 1 , wherein said EDA domain consists of an amino acid sequence selected from the group consisting of:
a) amino acid residues 2 to 91 of SEQ. ID. NO: 2; and
b) sequence SEQ. ID. NO: 4.
9. The proteinaceous vector according to claim 1 , wherein said EDA domain consists of an amino acid sequence selected from the group consisting of:
a) amino residues 2 to 57 of SEQ. ID. NO: 6; and
b) sequence SEQ. ID. NO: 8.
10. The proteinaceous vector according to claim 1 , wherein said molecule of interest is a parasitic antigen.
11. The proteinaceous vector according to claim 1 , wherein said molecule of interest is a fungal antigen.
12. The proteinaceous vector according to claim 1 , wherein the vector is capable of inducing a CTL response against the molecule of interest.
13. A pharmaceutical composition which comprises at least one acceptable pharmaceutical carrier, and at least one proteinaceous vector defined in claim 1 .
14. The pharmaceutical composition according to claim 13 , wherein the composition is a vaccine or an immunotherapeutic composition.
15. A method for stimulating an antigen-specific immune response in a human or animal subject in need thereof, comprising the administration to the subject of the proteinaceous vector bound to a molecule of interest according to claim 1 .
16. The method according to claim 15 , for the treatment or prophylaxis of a viral infectious disease, a fungal infectious disease, a parasitic infectious disease or a tumoral disease.
17. The method according to claim 16 , wherein the disease is a viral infectious disease.
18. The method according to claim 17 , wherein the viral infectious disease is hepatitis C.
19. The method according to claim 16 , wherein the disease is a tumoral disease.
20. The method according to claim 15 , wherein the EDA domain or the fragment of the EDA domain capable of binding TLR4 comprises a polypeptide whose amino acid sequence is selected from:
a) amino acids 2-91 of SEQ ID NO: 2;
b) SEQ ID NO: 4; and
c) a fragment of sequence a) or b).
21. The method according to claim 15 , wherein the EDA domain or the fragment of the EDA domain capable of binding TLR4 comprises a polypeptide whose amino acid sequence is selected from:
a) amino acids 2-57 of SEQ ID NO: 6;
b) SEQ ID NO: 8; and
c) a fragment of sequence a) or b).
22. The method according to claim 15 , wherein said antigen is a viral antigen from hepatitis C virus.
23. The method according to claim 15 , wherein said antigen is selected from a tumoral antigen and a tumoral antigenic determinant.
24. The method according to claim 22 , wherein the hepatitis C viral antigen is NS3 protein or an antigenic fragment of said protein.
25. The method according to claim 24 , wherein said amino acid sequence is SEQ ID NO: 10.