Trimeric OX40-immunoglobulin fusion protein and methods of use
Compositions including a trimeric OX-40 fusion protein are disclosed. Also disclosed are methods for enhancing the immune response of a mammal to an antigen by engaging the OX-40 receptor on the surface of T-cells involving administering to the mammal a composition comprising a trimeric OX-40 fusion protein and a pharmaceutically acceptable carrier.
1 . A fusion polypeptide comprising in an N-terminal to C-terminal direction:
an immunoglobulin domain;
a trimerization domain; and
a receptor binding domain, wherein the fusion polypeptide self-assembles into a trimeric fusion protein.
2 . The fusion polyeptide of claim 1 , wherein the receptor binding domain comprises a domain of a Tumor Necrosis Family ligand.
3 . The fusion polypeptide of claim 2 , wherein the receptor binding domain is an OX-40L receptor binding domain.
4 . The fusion polypeptide of claim 3 , wherein the fusion protein is capable of binding to the OX-40 receptor and stimulating at least one OX-40 mediated activity.
5 . The fusion polypeptide of claim 4 , OX-40 mediated activity is CD4 + T cell proliferation.
6 . The fusion polypeptide of claim 3 , wherein the OX-40 receptor binding domain comprises an extracellular domain of OX-40 ligand (OX-40L).
7 . The fusion polypeptide of claim 6 , wherein the OX-40 receptor binding domain comprises a polynucleotide sequence at least 95% identical to SEQ ID NO:2.
8 . The fusion polypeptide of claim 1 , wherein the trimerization domain comprises an isoleucine zipper domain.
9 . The fusion polypeptide of claim 1 , wherein the isoleucine zipper domain comprises a polynucleotide sequence at least 95% identical to SEQ ID NO:4.
10 . The fusion polypeptide of claim 1 , wherein the immunoglobulin domain comprises an Fc domain.
11 . The fusion polypeptide of claim 1 , comprising an amino acid sequence at least 95% identical to SEQ ID NO:8.
12 . The fusion polypeptide of claim 11 , wherein the polypeptide is encoded by a nucleic acid that hybridizes under highly stringent conditions to a nucleic acid with the polynucleotide sequence of SEQ ID NO:7.
13 . A fusion protein comprising a plurality of the fusion polypeptides of claim 1 .
14 . The fusion protein of claim 13 , consisting of three or six fusion polypeptides.
15 . The fusion protein of claim 13 , comprising an OX-40L fusion protein.
16 . A recombinant nucleic acid comprising a polynucleotide sequence that encodes the fusion polypeptide of claim 1 .
17 . The recombinant nucleic acid of claim 16 , the polynucleotide sequence comprising in a 5′ to 3′ direction:
a polynucleotide sequence encoding an immunoglobulin domain;
a polynucleotide sequence encoding a trimerization domain; and
a polynucleotide encoding a ligand receptor binding domain.
18 . The recombinant nucleic acid of claim 17 , wherein the polynucleotide encoding a ligand receptor binding domain encodes an OX-40L domain.
19 . A pharmaceutical composition comprising the fusion polypeptide of claim 1 , or a nucleic acid encoding the fusion polypeptide.
20 . A fusion polypeptide comprising in an N-terminal to C-terminal direction:
a dimerization domain;
a trimerization domain; and
an OX-40 receptor binding domain, wherein the fusion polypeptide self-assembles into a trimeric fusion protein.
21 . The fusion polypeptide of claim 20 , wherein the dimerization domain is an immunoglobulin domain.
22 . The fusion polypeptide of claim 21 , wherein the immunoglobulin domnain is an Fc domain.
23 . The fusion polypeptide of claim 20 , wherein the trimerization domain is an isoleucine zipper domain.
24 . The fusion polypeptide of claim 20 , wherein the trimeric fusion protein consists of three or six fusion polypeptides.
25 . A method of enhancing an immune response in a subject, the method comprising:
administering to a subject exposed to an antigen, a therapeutically effective amount of the fusion protein of claim 1 , thereby enhancing the immune response to the antigen by the subject.
26 . The method of claim 25 , wherein the subject is a human subject.
27 . The method of claim 25 , wherein the subject is exposed to the antigen prior to administering theusion protein.
28 . The method of claim 25 , wherein the antigen and the trimeric OX-40L fusion protein are administered at the same time.
29 . The method of claim 25 , wherein the fusion protein is administered by expressing a recombinant nucleic acid encoding an OX-40L fusion polypeptide, which fusion polypeptide assembles into the fusion protein in at least one cell of the subject.
30 . The method of claim 29 , wherein the recombinant nucleic acid encoding the OX-40L fusion polypeptide is introduced ex vivo into at least one cell, and the at least one cell comprising the recombinant nucleic acid is introduced into the subject.
31 . The method of claim 29 , comprising administering the fusion protein by introducing into the subject a viral or plasmid vector comprising the nucleic acid encoding an OX-40L fusion polypeptide, which fusion polypeptide assembles into the fusion protein.
32 . The method of claim 31 , wherein the viral vector is an adenovirus vector, a retrovirus vector or a herpesvirus vector.