MHC complexes and uses thereof
The present invention relates to novel complexes of major histocompability complex (MHC) molecules and uses of such complexes. In one aspect, the invention relates to loaded MHC complexes that include at least one MHC molecule with a peptide-binding groove and a presenting peptide non-covalently linked to the MHC protein. In another aspect, the invention features single chain MHC class II peptide fusion complexes with a presenting peptide covalently linked to the peptide binding grove of the complex. MHC complexes of the invention are useful for a variety of applications including: 1) in vitro screens for identification and isolation of peptides that modulate activity of selected T cells, including peptides that are T cell receptor antagonists and partial agonists, and 2) methods for suppressing or inducing an immune response in a mammal.
1. A single chain class II MHC molecule comprising:
a peptide-binding groove and
covalently linked in sequence: 1) a class II β chain, 2) a single chain linker, and 3) a class II α chain,
wherein the chain of both 1) and 3) lack a transmembrane domain; and further wherein the single chain class II MHC molecule is empty.
2. The MHC molecule of claim 1 , wherein the MHC molecule is soluble.
3. The MHC molecule of claim 1 , wherein the chain of 1) comprises a β1 domain and the chain of 3) comprises an α1 domain.
4. The MHC molecule of claim 1 , wherein the single chain linker is linked between the carboxyl terminus of the β chain and the amino terminus of the α chain.
5. The MHC molecule of claim 1 , wherein the β and α chains are each independently selected from the group consisting of IE, IA, DR, DQ and DP proteins.
6. The MHC molecule of claim 1 wherein the MHC molecule is modified to carry a detectable tag.
7. A multivalent MHC complex comprising two or more linked MHC molecules of claim 1 .
8. A MHC complex of claim 7 wherein the MHC molecules are linked to immunoglobulin domains.
9. A MHC complex of claim 7 wherein the MHC complex is modified to carry a detectable tag.
10. A single chain MHC class II-peptide complex comprising:
a peptide-binding groove;
covalently linked in sequence: 1) a class II β chain, 2) a single chain linker, and 3) a class II α chain, wherein the chain of both 1) and 3) lack a transmembrane domain; and
a presenting peptide being covalently linked to the MHC molecule and non-covalently bound to the peptide binding groove of the MHC molecule.
11. The MHC complex of claim 10 , wherein the complex is soluble.
12. The MHC complex of claim 10 , wherein the chains of 1) and 3) comprise a β1 domain and α1 domain, respectively.
13. The MHC complex of claim 10 , wherein the MHC class II molecule comprises the presenting peptide covalently linked to the β chain.
14. The MHC complex of claim 10 , wherein a presenting peptide linker sequence is interposed between the presenting peptide and the MHC molecule.
15. The MHC complex of claim 10 , wherein the β and α chains are each independently selected from the group consisting of IE, IA, DR, DQ and DP proteins.
16. The MHC complex of claim 10 , wherein the MHC molecule is modified to carry a detectable tag.
17. A multivalent MHC complex comprising two or more linked MHC molecules of claim 10 .
18. The MHC complex of claim 17 , wherein the MHC molecules are linked to immunoglobulin domains.
19. The MHC complex of claim 17 , wherein the MHC complex is modified to carry a detectable tag.
20. A loaded single chain MHC class II-peptide complex comprising:
a peptide-binding groove;
covalently linked in sequence: 1) a class II β chain, 2) a single chain linker, and 3) a class II α chain, wherein the chain of both 1) and 3) lack a transmembrane domain; and
a presenting peptide being non-covalently linked to the MHC molecule and non-covalently bound to the peptide binding groove of the MHC molecule,
wherein the loaded single chain MHC class II-peptide complex can be recognized by a CD4+ T cell.
21. The MHC complex of claim 20 , wherein the MHC molecule is soluble.
22. The MHC complex of claim 20 , wherein the chain of 1) comprises a β1 domain and the chain of 3) comprises an α1 domain.
23. The MHC complex of claim 20 , wherein the single chain linker is linked between the carboxyl terminus of the β chain and the amino terminus of the α chain.
24. The MHC complex of claim 20 , wherein the β and α chains are each independently selected from the group consisting of IE, IA, DR, DQ and DP proteins.
25. The MHC complex of claim 20 wherein the MHC molecule is modified to carry a detectable tag.
26. A multivalent MHC complex comprising two or more linked MHC complexes of claim 20 .
27. The multivalent MHC complex of claim 26 wherein the MHC complexes are linked to immunoglobulin domains.
28. A multivalent MHC complex of claim 26 wherein the MHC complex is modified to carry a detectable tag.