Recombinant MHC molecules useful for manipulation of antigen-specific T-cells
AssigneeOregon Health & Science University
InventorsGregory G. Burrows, Arthur A. Vandenbark
Life of this patent
Estimating the term…
Abstract
Two-domain MHC polypeptides useful for manipulation of antigen-specific T-cells are disclosed. These polypeptides include MHC class II-based molecules that comprise covalently linked β1 and α1 domains, and MHC class I-based molecules that comprise covalently linked α1 and α2 domains. These polypeptides may also include covalently linked antigenic determinants, toxic moieties, and/or detectable labels. The disclosed polypeptides can be used to target antigen-specific T-cells, and are useful, among other things, to detect and purify antigen-specific T-cells, to induce or activate T-cells, and to treat conditions mediated by antigen-specific T-cells.
Claims33
A purified polypeptide comprising, covalently linked first and second domains, wherein:
- (a) the first domain is a mammalian MHC class II β1 domain and the second domain is a mammalian MHC class II α1 domain;
- or
- (b) the first domain is a mammalian MHC class I α1 domain and the second domain is a mammalian MHC class I α2 domain, and wherein the polypeptide is not a whole MHC class I α chain;
- and wherein the amino terminus of the second domain is covalently linked to the carboxy terminus of the first domain.
A recombinant polypeptide comprising α1 and β1 domains of a mammalian molecule wherein the amino terminus of the α1 domain is covalently linked to the carboxy terminus of the β1 domain.
A polypeptide comprising covalently joined α1 and α2 domains of a mammalian MHC class I molecule wherein the amino terminus of the α2 domain is covalently linked to the carboxy terminus of the α1 domain, and wherein the polypeptide does not include an MHC class I α3 domain.
A recombinant nucleic acid molecule, comprising first, second and third regions represented by the formula Pr-B-A, wherein:
- Pr is a promoter sequence;
- B is a coding sequence that encodes a β1 domain of a mammalian MHC class II molecule;
- and
- A is a coding sequence that encodes an a 1 domain of a mammalian MHC class II molecule;
- wherein Pr is operably linked to B, and B and A comprise a single open reading frame.
A recombinant nucleic acid molecule, comprising first, second, third and fourth regions represented by the formula Pr-P-B-A, wherein:
- Pr is a promoter sequence;
- P is a coding sequence that encodes a peptide antigen;
- B is a coding sequence that encodes a β1 domain of a mammalian MHC class II molecule;
- and
- A is a coding sequence that encodes an α1 domain of a mammalian MHC class II molecule;
- wherein Pr is operably linked to P, and P, B and A comprise a single open reading frame.
A recombinant nucleic acid molecule, comprising first, second and third regions represented by the formula Pr-B-A, wherein:
- Pr is a promoter sequence;
- B is a coding sequence that encodes an α1 domain of a mammalian MHC class I molecule;
- and
- A is a coding sequence that encodes an α2 domain of a mammalian MHC class I molecule;
- wherein Pr is operably linked to B, and B and A comprise a single open reading frame, and wherein the open reading frame does not encode an α3 domain of a mammalian MHC class I molecule.
A recombinant nucleic acid molecule, comprising first, second, third and fourth regions represented by the formula Pr-P-B-A, wherein:
- Pr is a promoter sequence;
- P is a coding sequence that encodes a peptide antigen;
- B is a coding sequence that encodes an α1 domain of a mammalian MHC class I molecule;
- and
- A is a coding sequence that encodes an α2 domain of a mammalian MHC class I molecule;
- wherein Pr is operably linked to P, and P, B and A comprise a single open reading frame, and wherein the open reading frame does not encode an α3 domain of a mammalian MHC class I molecule.
A method for detecting or quantifying in a biological sample the presence of T-cells having a receptor specific for a specified antigen, comprising:
- combining the biological sample with a recombinant polypeptide comprising covalently linked α1 and β1 domains of a mammalian MHC, class II molecule wherein the carboxy terminus of the β1 domain is covalently linked to the amino terminus of the α1 domain, and further comprising the specified antigen bound in a peptide binding groove formed by said α1 and β1 domains;
- and
- detecting or quantifying the presence of specific binding of the recombinant polypeptide with said T-cells.
A method for separating T-cells having a receptor specific for a specified antigen from a mixture of cells, comprising:
- combining the cell mixture with a recombinant polypeptide comprising covalently linked α1 and β1I domains of a mammalian MHC class II molecule wherein the carboxy terminus of the β1 domain is covalently linked to the amino terminus of the α1 domain, and further comprising the specified antigen bound in a peptide binding groove formed by said α1 and β1 domains;
- and
- separating those cells bound to the recombinant polypeptide from unbound cells.
A method for detecting or quantifying in a biological sample the presence of T-cells having a receptor specific for a specified antigen, comprising:
- combining the biological sample with a polypeptide comprising covalently linked α1 and α2 domains of a mammalian MHC class I molecule wherein the carboxy terminus of the α1 domain is covalently linked to the amino terminus of the α2 domain, wherein the polypeptide does not include an α3 domain of a mammalian MHC class I molecule and wherein the polypeptide further comprises the specified antigen bound in a peptide binding groove formed by said α1 and α2 domains;
- and
- detecting or quantifying the presence of specific binding of the recombinant polypeptide with said T-cells.
A method for separating T-cells having a receptor specific for a specified antigen from a mixture of cells, comprising:
- combining the cell mixture with a polypeptide comprising covalently linked α1 and α2 domains of a mammalian MHC class I molecule wherein the carboxy terminus of the α1 domain is covalently linked to the amino terminus of the α2 domain, wherein the polypeptide does not include an α3 domain of a mammalian MHC class I molecule and wherein the polypeptide further comprises the specified antigen bound in a peptide binding groove formed by said α1 and α2 domains;
- and
- separating those cells bound to the recombinant polypeptide from unbound cells.
Dependencies are inferred from the claim text.