Product and process for T cell regulation

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 .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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 .alpha. and .beta.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.

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 .beta. chain and at least a portion of an MHC class II .alpha. 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 .alpha. and .beta. 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 .alpha. and .beta. 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.

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 .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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 .alpha. and .beta.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.

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 .beta. chain and at least a portion of an MHC class II .alpha. 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 .alpha. and .beta. 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 .alpha. and .beta. 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.

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 .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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 .alpha. and .beta.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.

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 .beta. chain and at least a portion of an MHC class II .alpha. 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 .alpha. and .beta. 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 .alpha. and .beta. 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.

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 .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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 .alpha. and .beta.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.

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 .beta. chain and at least a portion of an MHC class II .alpha. 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 .alpha. and .beta. 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.

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 .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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.

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 .beta. chain and at least a portion of an MHC class II .alpha. 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 .alpha. and .beta. 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.

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 .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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.

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 .beta. chain and at least a portion of an MHC class II .alpha. 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 .alpha. and .beta. 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 .alpha. and .beta. 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.

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 .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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 .alpha. and .beta.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.

A method for producing a Peptide-L-MHC.sub..alpha.+.beta. heterodimer, comprising:

• a) culturing a cell transformed with at least one nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC.sub..alpha.+.beta. heterodimer, comprising:
• (i) an MHC segment of said heterodimer comprising at least a portion of an MHC class II .beta. chain and at least a portion of an MHC class II .alpha. 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 .alpha. and .beta. 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..alpha.+.beta. heterodimer and said T cell receptor;
• wherein said antigenic peptide, linker and at least one of said MHC class II .alpha. and .beta. 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..alpha.+.beta. heterodimer;
• and
• b) recovering said Peptide-L-MHC.sub..alpha.+.beta. heterodimer.

A method for producing a Peptide-L-MHC.sub..alpha.+.beta.2m heterodimer, comprising:

• a) culturing a cell transformed with at least one nucleic acid molecule comprising a sequence encoding a Peptide-L-MHC.sub..alpha.+.beta.2m heterodimer, comprising:
• (i) an MHC segment of said heterodimer comprising at least a portion of an MHC class I .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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..alpha.+.beta.2m heterodimer and said T cell receptor;
• wherein said antigenic peptide, linker and at least one of said MHC class I .alpha. and .beta.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..alpha.+.beta.2m heterodimer;
• and
• b) recovering said Peptide-L-MHC.sub..alpha.+.beta.2m heterodimer.

A method for producing a Peptide-L-MHC.sub..alpha.+.beta. 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..alpha.+.beta. heterodimer comprising:
• (i) an MHC segment of said heterodimer comprising at least a portion of an MHC class II .alpha. 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 .beta. 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 .alpha. and .beta. 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..alpha.+.beta. heterodimer and said T cell receptor;
• wherein said antigenic peptide, linker and at least one of said MHC class II .alpha. and .beta. 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..alpha.+.beta. heterodimer anchored to the plasma membrane of said cell;
• and
• b) recovering said cell.

A method for producing a Peptide-L-MHC.sub..alpha.+.beta.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..alpha.+.beta.2m heterodimer comprising:
• (i) an MHC segment of said heterodimer comprising at least a portion of an MHC class I .beta.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 .alpha. 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 .alpha. and .beta.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..alpha.+.beta.2m heterodimer and said T cell receptor;
• wherein said antigenic peptide, linker and at least one of said MHC class I .alpha. and .beta.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..alpha.+.beta.2m heterodimer anchored to the plasma membrane of said cell;
• and
• b) recovering said cell.

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 .beta. chain and at least a portion of an MHC class II .alpha. 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 .alpha. and .beta. 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 .alpha. and .beta. 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.

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 .alpha. chain and at least a portion of an MHC class I .beta.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 .alpha. and .beta.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 .alpha. and .beta.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.