Product and process for T cell regulation

• US 5,820,866 A
• Filed: 03/04/1994
• Issued: 10/13/1998
• Est. Priority Date: 03/04/1994
• Status: Expired due to Term

• Current Assignee: National Jewish Center for Immunology and Respiratory Medicine
• Original Assignee: National Jewish Center for Immunology and Respiratory Medicine
• Inventors: Kappler, John W., Marrack, Philippa
• Primary Examiner(s): Cunningham, Thomas M.
• Application Number:
  US08/207,481
• Time in Patent Office: 1,684 Days
• Field of Search: 424/185.1, 424/193.1, 424/192.1, 530/300, 530/328, 530/350, 530/395, 530/868, 530/403, 435/69.7, 435/69.8, 435/69.3, 536/23.5, 536/23.4
• US Class Current: 424/192.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/30 Animal model comprising expression system for selective cell killing, e.g. toxins, enzyme dependent prodrug therapy using ganciclovir
• CPC Class Codes: A01K 2267/0325 Animal model for autoimmune diseases
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: C07K 14/70539 MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C07K 14/77 Ovalbumin
• CPC Class Codes: C07K 14/80 Cytochromes
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/8509 : for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2799/026 : where the vector is derived from a baculovirus

Abstract

The present invention relates to a product and process for regulating the activity of T cells using major histocompatibility complexes (MHC) stably linked to antigenic peptides. Disclosed is an antigenic peptide covalently linked to a major histocompatibility complex (MHC) protein by a novel linker, thereby enabling the formation of a stable peptide-MHC complex, alone or in combination with additional MHC protein chains, capable of being recognized by a T cell receptor (TCR). Also disclosed is a nucleic molecule having a sequence encoding a Peptide-L-MHC molecule comprising an antigenic peptide joined by a linker to an MHC segment. The invention is additionally directed to formulations comprising an antigenic peptide joined by a linker to an MHC segment anchored in a lipid-containing substrate. Pharmaceutical reagents are also disclosed which contain an antigenic peptide joined by a linker to an MHC segment combined with a suitable carrier that is capable of presenting the Peptide-L-MHC molecule so that it is capable of being recognized by a T cell receptor.

Claims

1. An isolated Peptide-L-MHC heterodimer comprising:

a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain;

b) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

c) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L- MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class II α and

β

chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide.

3. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said peptide is at least a portion of an antigen selected from the group consisting of autoantigens, infectious antigens, toxins, and allergens.

4. The Peptide-L-MHC heterodimer of claim 1, wherein said peptide is selected from the group consisting of:

Arg-Ala-Asp-Leu-Ile-Ala-Tyr-Leu-Lys-Gln-Ala-Thr-Lys (SEQ ID NO;

1), Val-His-Ala-Ala-His-Ala-Glu-Ile-Asn-GIu-Ala-Gly-Arg (SEQ ID NO;

2) and Ala-Ser-Phe-Glu-Ala-Gln-Gly-Ala-Leu-Ala-Asn- Iso!Ile-Ala-Val-Asp-Lys-Ala (SEQ ID NO;

5).

5. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said linker is processable.

6. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said linker is substantially neutral.

7. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said linker comprises residues selected from the group consisting of glycine, alanine, serine, valine, threonine and proline.

8. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said linker comprises Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser (SEQ ID NO:

7).

9. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said linker comprises Gly-Gly-Gly-Gly-Ser-Leu-Val-Pro-Arg-Gly-Ser-Gly-Gly-Gly-Gly-Ser (SEQ ID NO:

6).

10. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said Peptide-L-MHC heterodimer is secreted from a cell that produces said Peptide-L-MHC heterodimer.

11. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said Peptide-L-MHC heterodimer is anchored to an artificial or natural lipid-containing substrate.

12. The Peptide-L-MHC heterodimer of claim 11, wherein said lipid-containing substrate is selected from the group consisting of cells, cellular membranes, liposomes and micelles.

13. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said Peptide-L-MHC heterodimer is anchored to the plasma membrane of a mammalian cell.

14. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said nucleic acid molecule encodes a Peptide-L-MHC heterodimer further comprising at least one transmembrane segment which anchors said Peptide-L-MHC heterodimer to a lipid-containing substrate.

15. The Peptide-L-MHC heterodimer of claim 1 or 2, wherein said nucleic acid molecule is operatively linked to an expression vector to form a recombinant molecule.

16. The Peptide-L-MHC heterodimer of claim 15, wherein said Peptide-L-MHC recombinant molecule is transformed into a host cell to produce a recombinant cell which expresses said recombinant molecule.

17. The Peptide-L-MHC heterodimer of claim 16, wherein said host cell is selected from the group consisting of bacterial, fungal, insect and mammalian cells.

18. The Peptide-L-MHC heterodimer of claim 16, wherein said host cell stimulates a T cell response.

19. The Peptide-L-MHC heterodimer of claim 16, wherein said host cell does not stimulate a T cell response.

20. The Peptide-L-MHC heterodimer of claim 16, wherein said host cell is selected from the group consisting of antigen presenting cells, fibroblasts, red blood cells, pluripotent progenitor cells, epithelial cells and neural cells.

21. The Peptide-L-MHC heterodimer of claim 1 or 2 produced by a method comprising:

a) culturing a cell transformed with a nucleic acid molecule comprising a sequence encoding said Peptide-L-MHC heterodimer, said step of culturing being conducted to produce said Peptide-L-MHC heterodimer; and

b) recovering said Peptide-L-MHC heterodimer.

2. An isolated Peptide-L-MHC heterodimer comprising:

a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain;

b) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

c) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class I α and

β

2m chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide.

22. A formulation which suppresses T cell activity comprising a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain;

(b) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class II α and

β

chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

wherein said heterodimer is anchored to the plasma membrane of a cell that does not stimulate a T cell response.

24. The formulation of claim 22 or 23, wherein said cell is selected from the group consisting of red blood cells, fibroblasts, pluripotent progenitor cells, epithelial cells and neural cells.

23. A formulation which suppresses T cell activity comprising a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain;

(b) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class I α and

β

2m chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

wherein said heterodimer is anchored to the plasma membrane of a cell that does not stimulate a T cell response.

25. A formulation which stimulates T cell activity comprising a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain;

(b) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class II α and

β

chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

wherein said heterodimer is anchored to the plasma membrane of a cell that stimulates a T cell response.

27. The formulation of claim 25 or 26, wherein said cell that stimulates a T cell response is selected from the group consisting of macrophages, B cells and dendritic cells.

26. A formulation which stimulates T cell activity comprising a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain;

(b) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide linker and at least one of said MHC class I α and

β

2m chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

wherein said heterodimer is anchored to the plasma membrane of a cell that stimulates a T cell response.

28. A formulation which suppresses T cell activity comprising a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain;

(b) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class II α and

β

chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

wherein said heterodimer is secreted from a cell that produces said heterodimer.

29. A formulation which suppresses T cell activity comprising a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain;

(b) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class I α and

β

2m chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

wherein said heterodimer is secreted from a cell that produces said heterodimer.

30. A nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain;

(b) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said nucleic acid molecule comprises a nucleic acid sequence encoding a signal segment covalently attached to the N-terminus of said antigenic peptide.

32. The nucleic acid molecule of claim 30 or 31, wherein said nucleic acid molecule further comprises a transmembrane segment.

33. The nucleic acid molecule of claim 30 or 31, wherein said nucleic acid molecule is operatively linked to an expression vector to form a recombinant molecule.

31. A nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain;

(b) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said nucleic acid molecule comprises a nucleic acid sequence encoding a signal segment covalently attached to the N-terminus of said antigenic peptide.

34. A recombinant cell comprising a recombinant molecule comprising a nucleic acid molecule operatively linked to an expression vector, said nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC heterodimer comprising;

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain;

(b) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said nucleic acid molecule comprises a nucleic acid sequence encoding a signal segment covalently attached to the N-terminus of said antigenic peptide.

36. The cell of claim 34 or 35, wherein said cell expresses said Peptide-L-MHC heterodimer.

37. The cell of claim 34 or 35, wherein said cell is selected from the group consisting of a bacterial, fungal, insect and mammalian cells.

38. The cell of claim 34 or 35, wherein said cell is selected from the group consisting of a mammalian cell and an insect cell.

39. The cell of claim 34 or 35, wherein said cell stimulates a T cell response.

40. The cell of claim 34 or 35, wherein said cell does not stimulate a T cell response.

41. The cell of claim 34 or 35, wherein said cell is selected from the group consisting of an antigen presenting cell, a fibroblast, a red blood cell, a pluripotent progenitor cell, epithelial cells and neural cells.

42. The cell of claim 34 or 35, wherein said cell is used to treat a disease that at least in part is caused by abnormal stimulation or suppression of the immune system.

35. A recombinant cell comprising a recombinant molecule comprising a nucleic acid molecule operatively linked to an expression vector, said nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain;

(b) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said nucleic acid molecule comprises a nucleic acid sequence encoding a signal segment covalently attached to the N-terminus of said antigenic peptide.

43. A pharmaceutical reagent comprising an isolated Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain;

(b) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove;

(c) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class II α and

β

chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide.

45. The pharmaceutical reagent of claim 43 or 44, wherein said pharmaceutical reagent further comprises a suitable carrier that presents said Peptide-L-MHC heterodimer to said T cell receptor.

46. The pharmaceutical reagent of claim 45, wherein said carrier comprises artificial or natural lipid-containing membrane.

47. The pharmaceutical reagent of claim 45, wherein said carrier is selected from the group consisting of cellular membranes, liposomes and micelles.

48. The pharmaceutical reagent of claim 45, wherein said carrier comprises a mammalian cell.

49. The pharmaceutical reagent of claim 43 or 44, wherein said Peptide-L-MHC heterodimer is soluble.

44. A pharmaceutical reagent comprising an isolated Peptide-L-MHC heterodimer comprising:

(a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain;

(b) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

(c) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class I α and

β

2m chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide.

50. A method for producing a Peptide-L-MHC.sub.α

+β

heterodimer, comprising;

a) culturing a cell transformed with at least one nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC.sub.α

+β

heterodimer, comprising;

(i) an MHC segment of said heterodimer comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain which form a peptide binding groove;

(ii) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

(iii) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said peptide binding groove form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC.sub.α

+β

heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class II α and

β

chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

said step of culturing being conducted to produce said Peptide-L-MHC.sub.α

+β

heterodimer; and

b) recovering said Peptide-L-MHC.sub.α

+β

heterodimer.

51. A method for producing a Peptide-L-MHC.sub.α

+β

2m heterodimer, comprising;

a) culturing a cell transformed with at least one nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC.sub.α

+β

2m heterodimer, comprising;

(i) an MHC segment of said heterodimer comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain which form a peptide binding groove;

(ii) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

(iii) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said peptide binding groove form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC.sub.α

+β

2m heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class I α and

β

2m chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

said step of culturing being conducted to produce said Peptide-L-MHC.sub.α

+β

2m heterodimer; and

b) recovering said Peptide-L-MHC.sub.α

+β

2m heterodimer.

52. A method for producing a Peptide-L-MHC.sub.α

+β

heterodimer anchored to the plasma membrane of a cell comprising;

a) culturing a cell transformed with at least one nucleic acid molecule having a sequence encoding a Peptide-L-MHC.sub.α

+β

heterodimer comprising;

(i) an MHC segment of said heterodimer comprising at least a portion of an MHC class II α

chain protein having at least a portion of a transmembrane domain and a cytoplasmic domain and at least a portion of an MHC class II β

chain having at least a portion of a transmembrane domain and a cytoplasmic domain which form a peptide binding groove;

(ii) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

(iii) a linker comprising an amino acid sequence of at least about 15 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class II α and

β

chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering the interaction of said Peptide-L-MHC.sub.α

+β

heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class II α and

β

chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

said step of culturing being conducted to produce said Peptide-L-MHC.sub.α

+β

heterodimer anchored to the plasma membrane of said cell; and

b) recovering said cell.

53. A method for producing a Peptide-L-MHC.sub.α

+β

2m heterodimer anchored to the plasma membrane of a cell comprising;

a) culturing a cell transformed with at least one nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC.sub.α

+β

2m heterodimer comprising;

(i) an MHC segment of said heterodimer comprising at least a portion of an MHC class I β

2m protein having at least a portion of a transmembrane domain and a cytoplasmic domain and at least a portion of an MHC class I α

chain having at least a portion of a transmembrane domain and a cytoplasmic domain which form a peptide binding groove;

(ii) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

(iii) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering the interaction of said Peptide-L-MHC.sub.α

+β

2m heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class I α and

β

2m chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide;

said step of culturing being conducted to produce said Peptide-L-MHC.sub.α

+β

2m heterodimer anchored to the plasma membrane of said cell; and

b) recovering said cell.

54. A method to regulate an immune response comprising administering to an animal an effective amount of an isolated Peptide-L-MHC heterodimer comprising:

a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class II β

chain and at least a portion of an MHC class II α

chain;

b) an antigenic peptide comprising from about 5 to about 40 amino acid residues which binds to said peptide binding groove; and

c) a linker comprising an amino acid sequence of at least about 15 amino acid residues, said peptide being covalently linked to the N-terminus of one of said MHC class II α and

β

chains of said MHC segment by said linker, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class II α and

β

chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide.

56. The method of claim 54 or 55, wherein said immune response is stimulated by said heterodimer.

57. The method of claim 54 or 55, wherein said immune response is suppressed by said heterodimer.

55. A method to regulate an immune response comprising administering to an animal an effective amount of an isolated Peptide-L-MHC heterodimer comprising:

a) an MHC segment of said heterodimer which forms a peptide binding groove, comprising at least a portion of an MHC class I α

chain and at least a portion of an MHC class I β

2m chain;

b) an antigenic peptide comprising from about 8 to about 20 amino acid residues which binds to said peptide binding groove; and

c) a linker comprising an amino acid sequence of at least about 8 amino acid residues, wherein said peptide is covalently linked by said linker to the N-terminus of one of said MHC class I α and

β

2m chains, said linker facilitating the binding of said peptide to said peptide binding groove such that said peptide and said MHC segment form a conformation that is recognized by a T cell receptor without substantially hindering interaction between said Peptide-L-MHC heterodimer and said T cell receptor;

wherein said antigenic peptide, linker and at least one of said MHC class I α and

β

2m chains is encoded by a nucleic acid molecule, said nucleic acid molecule including a nucleic acid sequence encoding a signal segment attached to the N-terminus of said antigenic peptide.