Compositions and methods for the detection, quantitation and purification of antigen-specific T cells
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsJohn D. Altman, Michael G. McHeyzer-Williams, Mark M. Davis
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Abstract
T cells are specifically labeled according to their antigen receptor by binding of a multimeric binding complex. The complex is prepared with major histocompatibility complex protein subunits having a homogeneous population of peptides bound in the antigen presentation site. The multimeric MHC-antigen complex forms a stable structure with T cells, thereby allowing for the labeling, identification and separation of specific T cells.
Claims16
A method for detecting mammalian T cells according to the specificity of their antigen receptor, the method comprising:
- combining a suspension of said T cells and a multimeric binding complex having the formula:
- (.alpha.-.beta.-P).sub.n, wherein .alpha.-.beta.-P is an MHC peptide complex monomer
- .alpha. comprises a soluble form of an .alpha. chain of a class I or class II MHC protein,
- .beta. comprises a soluble form of (i) a .beta. chain of a class II MHC protein or (ii) .beta..sub.2 microglobulin for a class I MHC protein;
- and
- P comprises a substantially homogeneous peptide antigen bound in the groove formed by two membrane distal domains of (i) said .alpha. chain for a class I MHC protein or (ii) said .alpha. chain and said .beta. chains for a class II MHC protein;
- wherein n is greater than or equal to 2, and said multimeric binding complex is formed by binding said monomer to a multivalent entity through specific attachment sites on said .alpha. or said .beta. chain;
- detecting the presence of specific binding of said multimeric binding complex and said T cells;
- wherein said complex is bound to T cells comprising an antigen receptor specific for said peptide antigen presented by said .alpha. and said .beta. MHC chains.
A method for separation of mammalian T cells according to their antigen receptor, the method comprising:
- combining a suspension of said T cells and a multimeric binding complex having the formula:
- (.alpha.-.beta.-P).sub.n, wherein .alpha.-.beta.-P is an MHC peptide complex monomer,
- .alpha. comprises a soluble form of an .alpha. chain of a class I or class II MHC protein,
- .beta. comprises a soluble form of (i) a .beta. chain of a class II MHC protein or (ii) .beta..sub.2 microglobulin for a class I MHC protein;
- and
- P comprises a substantially homogeneous peptide antigen bound in the groove formed by two membrane by distal domains of (i) said .alpha. chain for a class I MHC protein or (ii) said .alpha. chain and said .beta. chains for a class II MHC protein;
- wherein n is greater than or equal to 2, and said multimeric binding complex is formed by binding said monomer to a multivalent entity through specific attachment sites on said .alpha. or said .beta. chain;
- separating cells bound to said complex from unbound cells;
- wherein said complex is bound to T cells comprising an antigen receptor specific for said peptide antigen presented by said .alpha. and said .beta. MHC chains.
A multimeric binding complex for labeling mammalian T cells according to their antigen receptor, having the formula:
- (.alpha.-.beta.-P).sub.n, wherein .alpha.-.beta.-P is an MHC peptide complex monomer,
- .alpha. comprises a soluble form of an .alpha. chain of a class I or class II MHC protein,
- .beta. comprises a soluble form of (i) a .beta. chain of a class II MHC protein or (ii) P2 microglobulin for a class I MHC protein;
- and
- P comprises a substantially homogeneous peptide antigen bound in the groove formed by two membrane distal domains of (i) said .alpha. chain for a class I MHC protein or (ii) said .alpha. chain and said .beta. chains for a class II MHC protein;
- wherein n is greater than or equal to 2, and said multimeric binding complex is formed by binding said monomer to a multivalent entity through specific attachment sites on said .alpha. said .beta. chain.
Dependencies are inferred from the claim text.