Dendritic cell co-stimulatory molecules
A novel costimulatory protein molecule, B7-DC, which is a member of the B7 family, is described as is DNA coding therefor and expression vectors comprising this DNA. B7-DC protein, fragments, fusion polypeptides/proteins and other functional derivatives, and transformed cells expressing B7-DC are useful in vaccine compositions and methods. Compositions and methods are disclosed for inducing potent T cell mediated responses that can be harnessed for anti-tumor and anti-viral immunity.
1. A fusion polypeptide comprising a first fusion partner consisting of all or part of a B7-DC protein extracellular domain which
(i) is fused directly to a second polypeptide or
(ii) is fused to a linker peptide sequence that is fused to the second polypeptide,
wherein the B7-DC protein has at least 70% sequence identity to SEQ ID NO:2.
2. A dimeric or trimeric fusion polypeptide which is a dimer or trimer of the fusion polypeptide of claim 1 .
3. A vaccine composition useful for inducing an immune response against an antigen associated with a pathogenic cell or microorganism, comprising
(a) the fusion protein of claim 1 ;
(b) the antigen; and
(c) a pharmaceutically and immunologically acceptable excipient or carrier.
4. The fusion protein of claim 1 , wherein the second polypeptide comprises one or more domains of an Ig heavy chain constant region.
5. The fusion protein of claim 4 , wherein the one or more domains of the Ig heavy chain comprise C H 2 or C H 3 regions of a human immunoglobulin Cγ1 chain.
6. The fusion protein of claim 1 , wherein the first fusion partner is the extracellular domain encoded by nucleotides 58-663 of SEQ ID NO:1.
7. The fusion protein of claim 1 , wherein the extracellular domain comprises amino acids from about position 20-221 of SEQ ID NO:2.
8. The fusion protein of claim 1 , wherein the fusion protein binds to Programmed Death-1 (PD-1).
9. The fusion protein of claim 1 , wherein the fusion protein co-stimulates T cells in the presence of an adequate stimulus to the T cell receptor to proliferate or to produce and secrete cytokines.
10. The fusion protein of claim 4 , wherein the fusion protein is a dimeric fusion protein formed by covalent bonding of Cys residues in C H regions of two Ig heavy chains.
11. The fusion protein of claim 10 wherein the Cys residues are the same Cys residues that are disulfide linked in dimerized Ig heavy chains.
12. A fusion polypeptide of the extracellular domain of murine or human B7-DC, wherein the B7-DC polypeptide is selectively expressed on dendritic cells as compared to activated macrophages, wherein murine B7-DC polypeptide has 32% homology to human B7-H1, wherein the B7-DC polypeptide comprises single IgV and IgC domains, wherein the B7-DC polypeptide comprises a single transmembrane domain, wherein the murine B7-DC polypeptide comprises an intracytoplasmic tail 4 amino acids in length, wherein the B7-DC polypeptide does not bind to CD28 or CTLA-4 and does not include the CD28/CTLA-4 binding motifs, and wherein the 137-DC polypeptide is capable of co-stimulating T cells, with a second polypeptide.
13. The fusion polypeptide of claim 12 wherein the second polypeptide comprises: (a) one or more domains of an Ig heavy chain constant region; (b) two C domains of an IgG heavy chain constant region; or (c) the hinge, C H 2 and C H 3 regions of a human immunoglobulin Cγ1 chain.
14. The fusion polypeptide of claim 12 wherein the fusion polypeptide binds to a binding partner molecule on T cells and co-stimulates the T cells.
15. The fusion polypeptide of claim 14 wherein the binding partner molecule is a receptor on T cells that is not CD28 or CTLA-4.
16. The fusion polypeptide of claim 12 , wherein the second polypeptide comprises one or more domains of an Ig heavy chain constant region.
17. A mature polypeptide comprising a first fusion partner and a second fusion partner, wherein the first fusion partner consists of an extracellular domain of a B7-DC protein comprising an IgV domain,
wherein the B7-DC protein has at least 70% sequence identity to SEQ ID NO:2.
18. A mature polypeptide comprising a first fusion partner and a second fusion partner, wherein the first fusion partner consists of an extracellular domain of B7-DC comprising amino acid residues 20-221 of SEQ ID NO:2.
19. The mature polypeptide of claim 17 , wherein the polypeptide co-stimulates T cells.
20. The fusion polypeptide of claim 1 wherein the extracellular domain comprises an amino acid sequence extending from the cysteine at position 42 of SEQ ID NO:2 to the cysteine at position 102 of SEQ ID NO:2.
21. The fusion polypeptide of claim 1 wherein the extracellular domain comprises an amino acid sequence extending from the cysteine at position 42 of SEQ ID NO:2 to the cysteine at position 192 of SEQ ID NO:2.
22. The fusion polypeptide of claim 1 wherein the extracellular domain comprises an IgV domain.
23. The fusion polypeptide of claim 1 wherein the extracellular domain comprises an IgV and an IgC domain.
24. The mature polypeptide of claim 17 wherein the extracellular domain of B7-DC comprises an IgV domain and an IgC domain.
25. The vaccine composition of claim 3 further comprising a general immunostimulatory agent or adjuvant.
26. A fusion polypeptide comprising a first fusion partner consisting of amino acids 20-221 of SEQ ID NO:2 and a second fusion partner consisting of the hinge, C H 2 and C H 3 regions of a human immunoglobulin Cγ1 chain.
27. The fusion polypeptide of claim 26 further comprising a linker between the first and second fusion partners.
28. The fusion polypeptide of claim 26 wherein the first fusion partner is directly linked to the second fusion partner.
29. The fusion polypeptide of claim 28 , wherein the fusion polypeptide is a dimeric fusion protein formed by covalent bonding of Cys residues in at least one C H region of a first second fusion partner with at least one C H region of a second fusion partner.
30. A pharmaceutical composition comprising the fusion polypeptide of any one of claims 26 to 29 .
31. The fusion polypeptide of claim 12 , wherein the fusion protein binds to Programmed Death-1 (PD-1).